Compositions of grapiprant and methods for using the same

ABSTRACT

The present disclosure provides a method for treating pain or inflammation in a non-human animal in need thereof. The method comprises administering to a non-human animal a pharmaceutical composition comprising a therapeutically effective amount of grapiprant. Also provided herein are pharmaceutical compositions for treating pain or inflammation in a non-human animal in need thereof. The pharmaceutical compositions comprise a therapeutically effective amount of grapiprant and an excipient, including flavorants.

CROSS-REFERENCE

This present application is continuation of U.S. application Ser. No.15/144,609 filed May 2, 2016, and entitled “Compositions of Grapiprantand Methods for Using the Same” which, is a divisional of U.S.application Ser. No. 14/639,898 filed Mar. 5, 2015, and entitled“Compositions of Grapiprant and Methods for Using the Same,” whichclaims priority to U.S. provisional application Ser. No. 62/089,713filed Dec. 9, 2014, and entitled “Compositions of Grapiprant and Methodsfor Using the Same,” the entire disclosures of which are incorporatedherein by reference.

TECHNICAL FIELD

The present disclosure generally relates to compositions of grapiprantand methods for using the same.

BACKGROUND

Grapiprant is a prostaglandin E2 subtype 4 (EP₄) receptor antagonistuseful in treating pain and inflammation. Formulations of grapiprant arevaluable because they alter the pharmacokinetic properties of the drugwithin an organism, such as the peak plasma concentration, timeachieved, the half-life, and the area under the plasma concentrationcurve (C_(max), T_(max), t_(1/2), and AUC, respectively). Anotherconsideration while formulating a pharmaceutical composition is thepalatability of the dosage form, which improves the animal's compliancein taking the medicine.

SUMMARY

Briefly, therefore, one aspect of the present disclosure encompasses amethod for treating pain or inflammation in a non-human animal in needthereof. The method comprises administering to a non-human animal apharmaceutical composition comprising a therapeutically effective amountof grapiprant. The pharmaceutical composition may be administeredorally. The administering may achieve a C_(max) of grapiprant of about375 ng/mL to about 10000 ng/mL at a T_(max) of about 0.4 to about 3.4hours, such as C_(max) of grapiprant of 750 about ng/mL to about 4000ng/mL, or of about 1300 ng/mL to about 4000 ng/mL. The C_(max) ofgrapiprant may also be achieved at a T_(max) of about 0.7 to about 1.7hours, such as a T_(max) of about 0.5 to about 1.0 hours. The grapiprantis administered at a dosage rate of about 1 to about 10 mg per kilogrambodyweight of the non-human animal per day (mg/kg/day), morespecifically a dosage rate of about 2 to about 4 mg/kg/day. Thepharmaceutical composition may be administered at least once daily, orat least twice daily, such as at least thrice daily. The pharmaceuticalcomposition may be administered until the cause of pain subsides, forexample from about 6 days to about 9 months. The non-human animal may bea companion animal, such as a dog, cat, or horse. In exemplaryembodiments, the pharmaceutical formulation may be administered twicedaily at a dosage of about 2 to about 4 mg/kg/day for about 9 to about21 days, and the administering may achieve a C_(max) of grapiprant ofabout 750 ng/mL to about 2200 ng/mL at a T_(max) of about 0.7 to about1.7 hours. In other embodiments, the pharmaceutical formulation may beadministered to the non-human animal at about 10 to about 18 hoursbefore surgery.

Another aspect of the disclosure provides a pharmaceutical compositionfor treating pain or inflammation in a non-human animal in need thereof.The pharmaceutical composition comprises a therapeutically effectiveamount of grapiprant and an excipient. The excipient may comprise one ormore selected from the group consisting of lactose, sodium starchglycolate, microcrystalline cellulose, colloidal silicon dioxide,magnesium stearate, copovidone, and poloxamer. In particularembodiments, the pharmaceutical composition may comprise about 5% toabout 15% grapiprant (w/w of the total composition), about 20% to about80% lactose (w/w of the total composition), about 15% to about 80%microcrystalline cellulose (w/w of the total composition), about 1% toabout 10% sodium starch glycolate (w/w of the total composition), about1% to about 10% copovidone (w/w of the total composition), about 0.5% toabout 3% magnesium stearate (w/w of the total composition), about 0.5%to about 4% poloxamer (w/w of the total composition), and about 0.1% toabout 1% colloidal silicon dioxide (w/w of the total composition). Thepharmaceutical may further comprise about 1% to about 30% flavorant (w/wof the total composition), such as about 5% to about 15% flavorant (w/wof the total composition).

The disclosure also provides a method for treating pain or inflammationin a non-human animal in need thereof, comprising orally administeringto the non-human animal a pharmaceutical composition comprising atherapeutically effective amount of grapiprant; wherein theadministering achieves a C_(max) of grapiprant of 675 ng/mL to 5000ng/mL within 4 hours after administration and a half-life of less than14 hours. The C_(max) may be achieved within 1 hour afteradministration. The pharmaceutical formulation may be administered oncedaily at a dosage rate of 3 mg/kg/day to 15 mg/kg/day for 28 days, forexample at a rate of 3 mg/kg/day, 9 mg/kg/day, or 15 mg/kg/day.

Additional embodiments and features are set forth in part in thedescription that follows, and in part will become apparent to thoseskilled in the art upon examination of the specification, or may belearned by the practice of the embodiments discussed herein. A furtherunderstanding of the nature and advantages of certain embodiments may berealized by reference to the remaining portions of the specification thedrawings, the chemical structures, and descriptions, which forms a partof this disclosure.

BRIEF DESCRIPTION OF DRAWINGS

In addition to the exemplary aspects and embodiments described above,further aspects and embodiments will become apparent by reference to thedrawings and by study of the following descriptions.

FIG. 1 depicts individual serum concentrations of grapiprant over timefor Group 1 dogs, as described in Example 3.

FIG. 2 depicts individual serum concentrations of grapiprant over timefor Group 2 dogs, as described in Example 3.

FIG. 3 depicts individual serum concentrations of grapiprant over timefor Group 3 dogs, as described in Example 3.

FIG. 4 depicts individual serum concentrations of grapiprant over timefor Group 4 dogs, as described in Example 3.

FIG. 5 depicts individual serum concentrations of grapiprant over timefor Group 5 dogs, as described in Example 3.

FIG. 6 depicts individual serum concentrations of grapiprant over timefor Group 6 dogs as described in Example 3.

FIG. 7 depicts the combined individual serum concentrations forgrapiprant over time for Groups 1-6 dogs, as described in Example 3 andindividually shown in FIGS. 1-6.

FIG. 8 depicts individual serum concentrations of grapiprant over timefor Group 1 male dogs tested with Formulation A34, as described inExample 4.

FIG. 9 depicts individual serum concentrations of grapiprant over timefor Group 1 female dogs tested with Formulation A34, as described inExample 4.

FIG. 10 depicts individual serum concentrations of grapiprant over timefor Group 2 male dogs tested with Formulation A27, as described inExample 4.

FIG. 11 depicts individual serum concentrations of grapiprant over timefor Group 2 female dogs tested with Formulation A27, as described inExample 4.

FIG. 12 depicts individual serum concentrations of grapiprant over timefor Group 3 male dogs tested with Formulation A29, as described inExample 4.

FIG. 13 depicts individual serum concentrations of grapiprant over timefor Group 3 female dogs tested with Formulation A29, as described inExample 4.

FIG. 14 depicts individual serum concentrations of grapiprant over timefor Group 4 male dogs tested with Formulation A31, as described inExample 4.

FIG. 15 depicts individual serum concentrations of grapiprant over timefor Group 4 female dogs tested with Formulation A31, as described inExample 4.

FIG. 16 depicts the mean serum concentrations of grapiprant over timefor Group 1 dogs grouped by male and female, as described in Example 4and individually shown at FIGS. 8 and 9.

FIG. 17 depicts the mean serum concentrations of grapiprant over timefor Group 2 dogs grouped by male and female, as described in Example 4and individually shown at FIGS. 10 and 11.

FIG. 18 depicts the mean serum concentrations of grapiprant over timefor Group 3 dogs grouped by male and female, as described in Example 4and individually shown at FIGS. 12 and 13.

FIG. 19 depicts the mean serum concentrations of grapiprant over timefor Group 4 dogs grouped by male and female, as described in Example 4and individually shown at FIGS. 14 and 15.

FIG. 20 depicts the mean serum concentrations of grapiprant over timefor Groups 1-4 dogs, as described in Example 4 and shown variously aboveat FIGS. 8-19.

DETAILED DESCRIPTION

Grapiprant is a prostaglandin E2 subtype 4 (EP₄) receptor antagonist.Grapiprant has a CAS registry number of 415903-37-6 and is also referredto variously as AT-001, CJ-023,423, RQ-7, RQ-00000007, MR10A7, AAT-007,N-{2-[4-(2-ethyl-4,6-dimethyl-1H-imidazo[4,5-c]pyridin-1-yl)phenyl]ethyl}-N′-[(4-methylphenyl)sulfonyl]urea,N-[[[2-[4-(2-ethyl-4,6-dimethyl-1H-imidazo[4,5-c]pyridin-1-yl)phenyl]ethyl]amino]carbonyl]-4-methyl-benenesulfonamide, or2-ethyl-4,6-dimethyl-3-(4(2-(((((4-methylphenyl)sulfonyl)amino)carbonyl) amino)ethyl)phenyl)-3H-imidazo[4,5-c]pyridine. The chemicalstructure and synthesis of grapiprant are described in WO 2002/032900and U.S. Pat. Nos. 6,710,054, 7,141,580, and 7,479,564, the disclosuresof which are all included herein by reference in their entireties.Grapiprant has the following chemical structure:

Without wishing to be bound by theory, prostaglandin E2 (PGE2) is apotent modulator involved in the pathogenesis of a variety of diseasessuch as inflammation, pain, arthritis, and cancer. PGE2 binds to atleast four subtypes of PGE receptor, designated EP₁, EP₂, EP₃, and EP₄.Molecular pharmacology studies have revealed that all subtypes are7-transmembrane spanning receptors that belong to the G-protein coupledreceptor super family. EP₁ activation stimulates the release ofintracellular calcium; EP₂ and EP₄ stimulation both activate adenylatecyclase but differ in their response to certain ligands; and EP₃stimulation inhibits adenylate cyclase via inhibitory G-proteins.

In vivo, grapiprant inhibits [³H]PGE binding to human, rat, and dog EP₄receptors with a K_(i) of 13±4 nM, 20±1 nM, and 24.1±2.7 nMrespectively. Grapiprant is highly selective for the EP₄ receptor overother human prostanoid receptor subtypes and inhibits PGE₂-evokedelevation in intracellular cAMP at the human and rat EP₄ receptors withpA2 of 8.3±0.03 and 8.2±0.2 nM, respectively. Oral administration ofgrapiprant significantly reduces thermal hyperalgesia induced byintraplantar injection of PGE₂ (ED₅₀=12.8 mg/kg). Grapiprant iseffective in models of acute and chronic inflammatory pain. Grapiprantsignificantly reduces mechanical hyperalgesia induced by carrageenanmodel and reverses complete Freund's adjuvant-induced chronicinflammatory pain response. Taken together, grapiprant is a potent andselective antagonist of human and rat EP₄ receptors, producesantihyperalgesic effects in animal models of inflammatory pain.

Grapiprant may exist as any of several polymorphs. The polymorphs differfrom each other with respect to their physical properties, spectraldata, stability, and methods of preparation. Some crystalline forms havealready been described, for example Form A, Form B, Form C, Form D, andForm G as described in U.S. Pat. No. 7,960,407, ethyl acetate solvateForm I and Form II as described in WO 2012/157288, and From X, Form X2,Form X3, and Form B4 as described in co-pending application entitled“Crystalline Forms of Grapiprant,” the disclosures of which areincorporated by reference in their entireties.

(I) Pharmaceutical Compositions

One aspect of the disclosure provides for a pharmaceutical compositionfor treating pain or inflammation in a non-human animal in need thereof,comprising a therapeutically effective amount of grapiprant and anexcipient.

Grapiprant may be included within the composition in one or moreconcentrations. In general, the concentration of grapiprant may rangefrom about 1% to about 30% (w/w) of the total composition; that is, theamount of grapiprant may be from about 1% to about 30% by weight inrelation to all components in the pharmaceutical composition, includingthe grapiprant and excipients. In various embodiments, the concentrationof grapiprant may be from about 1% to about 2%, from about 2% to about3%, from about 3% to about 4%, from about 4% to about 5%, from about 5%to about 6%, from about 6% to about 7%, from about 7% to about 8%, fromabout 8% to about 9%, from about 9% to about 10%, from about 10% toabout 11%, from about 11% to about 12%, from about 12% to about 13%,from about 13% to about 14%, from about 14% to about 15%, from about 15%to about 16%, from about 16% to about 17%, from about 17% to about 18%,from about 18% to about 19%, from about 19% to about 20%, from about 20%to about 21%, from about 21% to about 22%, from about 22% to about 23%,from about 23% to about 24%, from about 24% to about 25%, from about 25%to about 26%, from about 26% to about 27%, from about 27% to about 28%,from about 28% to about 29%, or from about 29% to about 30% (w/w) of thetotal composition.

In exemplary embodiments, the concentration of grapiprant may be fromabout 5% to about 15% (w/w) of the total composition. In one embodiment,the concentration of grapiprant may less than about 30% (w/w) of thetotal composition. In another embodiment, the concentration ofgrapiprant may be more than about 1% (w/w) of the total composition.

In some embodiments, grapiprant may be provided in a pharmaceuticalcomposition at a concentration suitable to alleviate the pain orinflammation in a non-human animal. The grapiprant may be provided at aconcentration suitable to alleviate pain or inflammation in thenon-human animal in need thereof for about 6 days, for about 7 days, forabout 8 days, for about 9 days, for about 10 days, for about 11 days,for about 12 days, for about 13 days, for about 14 days, for about 15days, for about 16 days, for about 17 days, for about 18 days, for about19 days, for about 20 days, for about 21 days, for about 22 days, forabout 23 days, for about 24 days, for about 25 days, for about 26 days,for about 27 days, or for about 28 days.

In exemplary embodiments, the grapiprant may be provided at aconcentration suitable to alleviate pain or inflammation in thenon-human animal in need thereof for about 9 days to about 21 days. Inother exemplary embodiments, the grapiprant may be provided at aconcentration suitable to alleviate pain or inflammation in thenon-human animal in need thereof for about 12 days to about 14 days. Insome embodiments, the grapiprant may be provided at a concentrationsuitable to alleviate pain or inflammation in the non-human animal inneed thereof for about 1 month, about 2 months, about 3 months, about 4months, about 5 months, about 6 months, about 7 months, about 8 months,or about 9 months, or longer. In one embodiment, the grapiprant may beprovided at a concentration suitable to alleviate pain or inflammationin the non-human animal in need thereof for less than about 28 days. Inanother embodiment, the grapiprant may be provided at a concentrationsuitable to alleviate pain or inflammation in the non-human animal inneed thereof for at least about 9 days.

In other embodiments, grapiprant may be at least partially dissolved inan aqueous solvent (e.g., deionized and/or purified water). In someother embodiments, the grapiprant may be formulated as a suspension. Theconcentration of grapiprant within the composition may be at leastpartially dependent upon the route of administration and/or the numberof times in a pre-determined time period the composition is administeredto a non-human animal. For example, one or more compositions may bedesigned for injectable administration. As a result, the grapiprantwithin the composition may be delivered directly to the circulatorysystem (e.g., via intravenous administration), thereby circumventing theneed for absorption in the alimentary canal. Accordingly, greateramounts of grapiprant may reach the desired targets relative to oralformulations, leading to a lower necessary concentration of grapiprantin a sterile injectable version.

In other embodiments, the pharmaceutical composition may be orallyadministered one or more times per day, such as at least twice daily orat least thrice daily. For example, the pharmaceutical composition maybe administered as a solution, a suspension, a solid, or a viscousliquid formulation. Correspondingly, the greater number of times per daythe composition is administered to the non-human animal, a lesser theamount of grapiprant may produce the target result. In exemplaryembodiments, the pharmaceutical composition may be formulated for oraladministration, such as an oral solution or an oral suspension or anoral gel.

A variety of excipients commonly used in pharmaceutical formulations maybe selected on the basis of several criteria such as, for example, thedesired dosage form and the release profile properties of the dosageform. Non-limiting examples of suitable excipients include an agentselected from the group comprising a binder, a filler, anon-effervescent disintegrant, an effervescent disintegrant, apreservative, a diluent, a flavoring agent, a sweetener, a lubricant, anoral dispersing agent, a coloring agent, a taste masking agent, a pHmodifier, a stabilizer, a compaction agent, and combinations of any ofthese agents.

In one embodiment, the excipient may be a binder, which holds thepharmaceutical composition together until administration. Suitablebinders include starches, pregelatinized starches, gelatin,polyvinylpyrrolidone, cellulose, methylcellulose, sodium carboxymethylcellulose, ethylcellulose, polyacrylamides, polyvinyloxoazolidone,polyvinyl alcohols, C₁₂-C₁₈ fatty acid alcohol, polyethylene glycol,polyols, saccharides, oligosaccharides, polypeptides, peptides, andcombinations thereof.

In another embodiment, the excipient may be a filler, which adds bulk tothe pharmaceutical composition for easier handling and more accuratedosing. Suitable fillers include carbohydrates, inorganic compounds, andpolyvinylpyrrolidone. By way of non-limiting example, the filler may becalcium sulfate, e.g. both di- and tri-basic calcium sulfate; starch,calcium carbonate, magnesium carbonate, microcrystalline cellulose,dibasic calcium phosphate, magnesium oxide, calcium silicate, talc,modified starches, lactose, sucrose, mannitol, and sorbitol.

The excipient may be a non-effervescent disintegrant, which allows thepharmaceutical composition to more easily dissolve after administrationwithout evolving gas. Suitable examples of non-effervescentdisintegrants include starches (such as corn starch, potato starch, andthe like), pregelatinized and modified starches thereof, sweeteners,clays (such as bentonite), microcrystalline cellulose, alginates, sodiumstarch glycolate, and gums (such as agar, guar, locust bean, karaya,pecitin, and tragacanth).

In another embodiment, the excipient may be an effervescentdisintegrant, which allows the pharmaceutical composition to more easilydissolve during administration while evolving gas. By way ofnon-limiting example, suitable effervescent disintegrants include sodiumbicarbonate in combination with citric acid, and sodium bicarbonate incombination with tartaric acid.

The excipient may comprise a preservative, which increases the stabilityand storage lifetime of the pharmaceutical composition, particularlydelaying unwanted degradation of the active ingredient. Suitableexamples of preservatives include antioxidants (such as alpha-tocopherolor ascorbate) and antimicrobials (such as parabens, chlorobutanol orphenol). In other embodiments, an antioxidant such as butylatedhydroxytoluene (BHT) or butylated hydroxyanisole (BHA) may be used.

The excipient may include a diluent, which diminishes the relativeconcentrations of other components within the pharmaceuticalcomposition. Diluents suitable for use include pharmaceuticallyacceptable saccharides such as sucrose, dextrose, lactose,microcrystalline cellulose, fructose, xylitol, and sorbitol; polyhydricalcohols; starches; pre-manufactured direct compression diluents; andmixtures of any of the foregoing.

The excipient may comprise a surfactant, which alters the solubilityparameters of the other components within the pharmaceuticalcomposition. In various embodiments, the surfactant may be an alkylarylpolyether alcohol, such as Triton™ X-100, Surfonic™ N-100(nonoxaynol-10), or Witconol™ NP-100; or a poloxamer, such as Pluronic™,Synperonic™, or Kolliphor™. Other suitable examples of surfactantsinclude, for example, 2-acrylamido-2-methylpropane sulfonic acid, alkylpolyglycoside, ammonium perfluorononanoate, benzalkonium chloride (BAC),benzethonium chloride (BZT), 5-bromo-5-nitro-1,3-dioxane, cetyltrimethylammonium bromide (CTAB, hexadecyltrimehtylammonium bromide,cetyl trimethylammonium chloride), cetylpridinium chloride (CPC),cyclohexyl-1-hexyl-maltopyranoside, decylmaltopyranoside, decylpolyglucose, dimethyldioctadecylammonium chloride,dioctadecyldimethylammmonium bromide (DODAB),dipalmitoylphosphatidylcholine, lauryldimethylamine oxide,dodecylmaltopyranoside, magnesium laureth sulfate polyethoxylated tallowamine (POEA), octenidine dihydrochloride, octylphenoxypolyethoxyethanol(Igepal™ CA-630), octylthioglucopyranoside (OTG), ox gall, sodiumnonanoyloxybenzensulfonate, sorbitan monolaurate, surfactin, andthonozonium bromide. In exemplary embodiments, the surfactant may be apoloxamer or sodium lauryl sulfate.

The excipient may be a lubricant, which allows easier removal of thepharmaceutical composition from molds during manufacture and may aidadministration of the pharmaceutical composition. Suitable non-limitingexamples of lubricants include magnesium stearate, calcium stearate,zinc stearate, hydrogenated vegetable oils, sterotex, polyoxyethylenemonostearate, talc, polyethyleneglycol, sodium benzoate, sodium laurylsulfate, magnesium lauryl sulfate, and light mineral oil.

The excipient may be a dispersion enhancer, which aids dispersion of thecomponents of the pharmaceutical composition within the subject afteradministration. Suitable dispersants may include starch, alginic acid,polyvinylpyrrolidones, guar gum, kaolin, bentonite, purified woodcellulose, sodium starch glycolate, isomorphous silicate, andmicrocrystalline cellulose.

Depending upon the embodiment, it may be desirable to provide a coloringagent, which aids visualization and identification of the pharmaceuticalcomposition. Suitable color additives include food, drug and cosmeticcolors (FD&C), drug and cosmetic colors (D&C), or external drug andcosmetic colors (Ext. D&C). These colors or dyes, along with theircorresponding lakes, and certain natural and derived colorants may besuitable for use in the present disclosure depending on the embodiment.

In various embodiments, the excipient may include a pH modifier, whichmay alter the solubility profile and bioavailability parameters ofcomponents within the pharmaceutical composition. In certainembodiments, the pH modifier may include sodium carbonate or sodiumbicarbonate.

The weight fraction of the excipient or combination of excipients in thepharmaceutical composition may be about 98% or less, about 95% or less,about 90% or less, about 85% or less, about 80% or less, about 75% orless, about 70% or less, about 65% or less, about 60% or less, about 55%or less, about 50% or less, about 45% or less, about 40% or less, about35% or less, about 30% or less, about 25% or less, about 20% or less,about 15% or less, about 10% or less, about 5% or less, about 2%, orabout 1% or less of the total weight of the pharmaceutical composition.

In particular embodiments, the excipient may comprise one or moreselected from the group consisting of lactose, sodium starch glycolate,microcrystalline cellulose, colloidal silicon dioxide, magnesiumstearate, copovidone, surfactant, poloxamer, and sodium laurel sulfate.

The concentration of lactose within the pharmaceutical composition mayvary. In general, the concentration of lactose may range from about 10%to about 95% (w/w) of the total composition. In various embodiments, theconcentration of lactose may be from about 10% to about 15%, from about15% to about 20%, from about 20% to about 25%, from about 25% to about30%, from about 30% to about 35%, from about 35% to about 40%, fromabout 40% to about 45%, from about 45% to about 50%, from about 50% toabout 55%, from about 55% to about 60%, from about 60% to about 65%,from about 65% to about 70%, from about 70% to about 75%, from about 75%to about 80%, from about 80% to about 85%, from about 85% to about 90%,or from about 90% to about 95% (w/w) of the total composition. Inexemplary embodiments, the concentration of lactose may be from about20% to about 80% (w/w) of the total composition.

The concentration of microcrystalline cellulose within thepharmaceutical composition may vary. In general, the concentration ofmicrocrystalline cellulose may range from about 5% to about 95% (w/w) ofthe total composition. In various embodiments, the concentration ofmicrocrystalline cellulose may be from about 5% to about 10%, from about10% to about 15%, from about 15% to about 20%, from about 20% to about25%, from about 25% to about 30%, from about 30% to about 35%, fromabout 35% to about 40%, from about 40% to about 45%, from about 45% toabout 50%, from about 50% to about 55%, from about 55% to about 60%,from about 60% to about 65%, from about 65% to about 70%, from about 70%to about 75%, from about 75% to about 80%, from about 80% to about 85%,from about 85% to about 90%, or from about 90% to about 95% (w/w) of thetotal composition. In exemplary embodiments, the concentration ofmicrocrystalline cellulose may be from about 15% to about 80% (w/w) ofthe total composition.

The concentration of sodium starch glycolate within the pharmaceuticalcomposition may vary. In general, the concentration of sodium starchglycolate may range from about 1% to about 20% (w/w) of the totalcomposition. In various embodiments, the concentration of sodium starchglycolate may be from about 1% to about 2%, from about 2% to about 3%,from about 3% to about 4%, from about 4% to about 5%, from about 5% toabout 6%, from about 6% to about 7%, from about 7% to about 8%, fromabout 8% to about 9%, from about 9% to about 10%, from about 10% toabout 11%, from about 11% to about 12%, from about 12% to about 13%,from about 13% to about 14%, from about 14% to about 15%, from about 15%to about 16%, from about 16% to about 17%, from about 17% to about 18%,from about 18% to about 19%, or from about 19% to about 20% (w/w) of thetotal composition. In exemplary embodiments, the concentration of sodiumstarch glycolate may be from about 1% to about 10% (w/w) of the totalcomposition.

The concentration of copovidone within the pharmaceutical compositionmay vary. In general, the concentration of copovidone may range fromabout 1% to about 20% (w/w) of the total composition. In variousembodiments, the concentration of copovidone may be from about 1% toabout 2%, from about 2% to about 3%, from about 3% to about 4%, fromabout 4% to about 5%, from about 5% to about 6%, from about 6% to about7%, from about 7% to about 8%, from about 8% to about 9%, from about 9%to about 10%, from about 10% to about 11%, from about 11% to about 12%,from about 12% to about 13%, from about 13% to about 14%, from about 14%to about 15%, from about 15% to about 16%, from about 16% to about 17%,from about 17% to about 18%, from about 18% to about 19%, or from about19% to about 20% (w/w) of the total composition. In exemplaryembodiments, the concentration of copovidone may be from about 1% toabout 10% (w/w) of the total composition.

The concentration of magnesium stearate within the pharmaceuticalcomposition may vary. In general, the concentration of magnesiumstearate may range from about 0.1% to about 5% (w/w) of the totalcomposition. In various embodiments, the concentration of magnesiumstearate may be from about 0.1% to about 0.5%, from about 0.5% to about1%, from about 1% to about 1.5%, from about 1.5% to about 2%, from about2% to about 2.5%, from about 2.5% to about 3%, from about 3% to about3.5%, from about 3.5% to about 4%, from about 4% to about 4.5%, or fromabout 4.5% to about 5% (w/w) of the total composition. In exemplaryembodiments, the concentration of magnesium stearate may be from about0.5% to about 3% (w/w) of the total composition.

The concentration of surfactant within the pharmaceutical compositionmay vary. In general, the concentration of surfactant may range fromabout 0.1% to about 5% (w/w) of the total composition. In variousembodiments, the concentration of surfactant may be from about 0.1% toabout 0.5%, from about 0.5% to about 1%, from about 1% to about 1.5%,from about 1.5% to about 2%, from about 2% to about 2.5%, from about2.5% to about 3%, from about 3% to about 3.5%, from about 3.5% to about4%, from about 4% to about 4.5%, or from about 4.5% to about 5% (w/w) ofthe total composition. In exemplary embodiments, the concentration ofsurfactant may be from about 0.5% to about 4% (w/w) of the totalcomposition.

The concentration of colloidal silicon dioxide within the pharmaceuticalcomposition may vary. In general, the concentration of colloidal silicondioxide may range from about 0.1% to about 2% (w/w) of the totalcomposition. In various embodiments, the concentration of colloidalsilicon dioxide may be from about 0.1% to about 0.2%, from about 0.2% toabout 0.3%, from about 0.3% to about 0.4%, from about 0.4% to about0.5%, from about 0.5% to about 0.6%, from about 0.6% to about 0.7%, fromabout 0.7% to about 0.8%, from about 0.8% to about 0.9%, from about 0.9%to about 1.0%, from about 1.0% to about 1.1%, from about 1.1% to about1.2%, from about 1.2% to about 1.3%, from about 1.3% to about 1.4%, fromabout 1.4% to about 1.5%, from about 1.5% to about 1.6%, from about 1.6%to about 1.7%, from about 1.7% to about 1.8%, from about 1.8% to about1.9%, or from about 1.9% to about 2.0% (w/w) of the total composition.In exemplary embodiments, the concentration of colloidal silicon dioxidemay be from about 0.5% to about 1% (w/w) of the total composition.

In an exemplary embodiment, the pharmaceutical composition may compriseabout 5% to about 15% grapiprant (w/w), about 20% to about 80% lactose(w/w), about 15% to about 80% microcrystalline cellulose (w/w), about 1%to about 10% sodium starch glycolate (w/w), about 1% to about 10%copovidone (w/w), about 0.5% to about 3% magnesium stearate (w/w), about0.5% to about 4% surfactant (w/w), and about 0.1% to about 1% colloidalsilicon dioxide (w/w).

(i) Flavorants

It may be necessary to add one or more additional compounds to thepharmaceutical composition in order to increase palatability. Theexcipient may include a flavorant that may act as a flavoring agentand/or as a flavor-masking agent. In some embodiments, the flavorant maycomprise one or more of a sweetening agent, a savory agent (i.e., anagent that imbues the pharmaceutical composition with a salty flavor), abittering agent, and a souring agent. Flavorants may be chosen fromsynthetic flavor oils and flavoring aromatics and/or natural oils,extracts from plants, leaves, flowers, fruits, and combinations thereof.By way of example, these may include cinnamon oils, oil of wintergreen,peppermint oils, clover oil, hay oil, anise oil, eucalyptus, vanilla,citrus oils (such as lemon oil, orange oil, grape and grapefruit oil),and fruit essences (such as apple, peach, pear, strawberry, raspberry,cherry, plum, pineapple, and apricot). In some embodiments, theflavoring agents and/or flavor-masking agents may comprise avanilla-comprising composition, for example ethylvanillin, vanillin-RHD,vanillin-Merck, vanilla-TG-old, and suitable solvents (e.g., ethanoland/or water). In other embodiments, other flavorants may be added thatconfer other flavors on the composition, such as banana, pork liver,beef, etc.

In some embodiments, the flavoring agents and/or flavor-masking agentscan comprise a vanilla-comprising composition, such as, but not limitedto ethyl vanillin, vanillin (vanillin-RHD), natural vanilla flavor(vanillin-Merck), nature-identical vanilla flavor (vanilla-TG-old), andsuitable solvents (e.g., ethanol and/or water).

In other embodiments, the flavoring agents and/or flavor-masking agentscan comprise one or more selected from chicken, bacon, beef, pork,liver, fish, honey, caramel, and banana.

In another embodiment, the excipient may include a sweetener. By way ofnon-limiting example, the sweetener may be selected from glucose (cornsyrup), dextrose, invert sugar, fructose, and mixtures thereof (when notused as a carrier); saccharin and its various salts such as the sodiumsalt; dipeptide sweeteners such as aspartame; dihydrochalcone compounds,glycyrrhizin; stevia-derived sweeteners; chloro derivatives of sucrosesuch as sucralose; sugar alcohols such as sorbitol, mannitol, xylitol,and the like. Also contemplated are hydrogenated starch hydroly satesand the synthetic sweetener3,6-dihydro-6-methyl-1,2,3-oxathiazin-4-one-2,2-dioxide, particularlythe potassium salt (acesulfame-K), and sodium and calcium salts thereof.In particular embodiments, the pharmaceutical composition may beformulated for oral administration and include one or more of thefollowing flavorant (e.g., sweetening agents): sucralose, MagnaSweet®,Di-Pac® compressible sugar (i.e., a 97:3 mixture of sucrose andmaltodextrin), Thaumatin T200X, Talin-Pure, OptisweetSD, stevia extractrebaudioside A, and/or neotame.

In some embodiments, the pharmaceutical composition that may beformulated for oral administration can include one or more of thefollowing flavoring agents and/or flavor-masking agents (e.g.,sweetening agents): sucralose; a dispersion of licorice, licoricederivatives, and licorice extract (glycyrrhizic acid/monoammoniumglycyrrhizinate); MagnaSweet®; a blend of sodium saccharin andneohesperidin dihydrochalcone (Optisweet™ SD), 97:3 (w/w) mixture ofsucrose and maltodextrin (Di-Pac®), thaumatin 7% (sweetener) blendedwith an inactive maltodextrin (Thaumatin T200X), pure thaumatin(Talin-Pure), stevia extract rebaudioside A (steviol glycosides),neotame, and/or polyols (sugar alcohols), such as sorbitol, maltitol,isomalt, xylitol, and glycerin.

As used herein “MagnaSweet®” refers to a composition consistingessentially of one or more sweeteners selected from the group consistingof glycyrrhizic acid (GA), monoammonium glycyrrhizinate (MAG),rebaudioside A, and glycerin. In some embodiments, the MagnaSweet®consists essentially of glycyrrhizic acid (GA), monoammoniumglycyrrhizinate (MAG), rebaudioside A, and glycerin. In otherembodiments, the MagnaSweet® consists essentially of glycyrrhizic acid(GA), monoammonium glycyrrhizinate (MAG), and glycerin. In someembodiments, the MagnaSweet® comprises from about 0.5% to about 25%GA/MAG, from about 0% to about 15% rebaudioside A, and from about 75% toabout 99.5% glycerin. In other embodiments, the MagnaSweet® comprisesfrom about 1.5% to about 17% GA/MAG, from about 0% to about 7.5%rebaudioside A, and from about 83% to about 91% glycerin. In exemplaryembodiments, the MagnaSweet® comprises about 1.5% GA/MAG, about 7.5%rebaudioside A, and about 91% glycerin. In other exemplary embodiments,the MagnaSweet® comprises about 9% GA/MAG and about 91% glycerin. Inanother exemplary embodiment, the MagnaSweet® comprises about 17% GA/MAGand about 83% glycerin.

In particular, some sugar-containing sweeteners, such assaccharose-containing materials, sucrose, glucose, fructose, andmaltodextrin, may at least partially degrade the capromorelin within thecomposition. Accordingly, large concentrations of some sugar-containingsweeteners should be avoided.

In exemplary embodiments, the flavoring agents or masking agents cancomprise at least one of thaumatin, sucralose, neotame, sodiumsaccharain, neohesperidin dihydrochalcone, rebaudioside A, steviolglycosilde, licorice, glycyrrhizic acid, monoammonium glycyrrihizinate,sucrose, glucose, fructose, maltodextrin, sorbitol, maltitol, isomalt,glycerol, and a vanilla-comprising composition.

The excipient may include a taste-masking agent. Taste-masking materialsinclude cellulose hydroxypropyl ethers (HPC); low-substituted cellulosehydroxypropyl ethers (L-HPC); cellulose hydroxypropyl methyl ethers(HPMC); methylcellulose polymers and mixtures thereof polyvinyl alcohol(PVA); hydroxyethylcelluloses; carboxymethylcelluloses and saltsthereof; polyvinyl alcohol and polyethylene glycol co-polymers;monoglycerides or triglycerides; polyethylene glycols; acrylic polymers;mixtures of acrylic polymers with cellulose ethers; cellulose acetatephthalate; and combinations thereof.

In some embodiments, the flavorant may comprise a percent weight perfinal volume of the pharmaceutical composition form about 50% to about0.001%, depending on the agent selected, such as from about 40% to about0.01%, from about 30% to about 0.01%, from about 1% to about 30%, orfrom about 5% to about 15%. As previously mentioned, the pharmaceuticalcomposition may include more than one flavorant.

(ii) Dosage Form

The pharmaceutical compositions detailed herein may be manufactured inone or several dosage forms. Suitable dosage forms include tablets,including suspension tablets, chewable tablets, effervescent tablets orcaplets; pills; powders such as a sterile packaged powder, a dispensablepowder, and an effervescent powder; capsules including both soft or hardgelatin capsules such as HPMC capsules; lozenges; a sachet; a sprinkle;a reconstitutable powder or shake; a troche; pellets such as sublingualor buccal pellets; granules; liquids for oral or parenteraladministration; suspensions; emulsions; semisolids; or gels. Othersuitable dosage forms include transdermal systems or patches. Thetransdermal system may be a matrix system, a reservoir system, or asystem without rate-controlling membranes.

The dosage forms may be manufactured using conventional pharmacologicaltechniques. Conventional pharmacological techniques include, forexample, one or a combination of methods: (1) dry mixing, (2) directcompression, (3) milling, (4) dry or non-aqueous granulation, (5) wetgranulation, or (6) fusion. See, e.g., Lachman et al., The Theory andPractice of Industrial Pharmacy (1986). Other methods include, e.g.,prilling, spray drying, pan coating, melt granulation, granulation,wurster coating, tangential coating, top spraying, extruding,coacervation and the like.

The amount of active ingredient that is administered to a subject canand will vary depending upon a variety of factors such as the age andoverall health of the subject, and the particular mode ofadministration. Those skilled in the art will appreciate that dosagesmay also be determined with guidance from Goodman & Gilman's ThePharmacological Basis of Therapeutics, Tenth Edition (2001), AppendixII, pp. 475-493, and the Physicians' Desk Reference (PDR).

(II) Methods for Treatment Using a Grapiprant Composition

Another aspect of the present disclosure encompasses a method fortreating pain or inflammation in a non-human animal in need thereof. Themethod comprises administering to the non-human animal a pharmaceuticalcomposition comprising a therapeutically effective amount of grapiprant.

Dosage of grapiprant may range from about 0.01 milligrams of grapiprantper kilogram of bodyweight of the non-human animal (“mg/kg”) to about 75mg/kg, such as about 0.1 mg/kg to about 7.5 mg/kg. In some embodiments,the grapiprant dosage may range from about 0.75 mg/kg to about 6 mg/kg.In some embodiments, the grapiprant dosage may range from about 6 mg/kgto about 50 mg/kg. In other embodiments, the grapiprant dosage may be atleast about 0.1 mg/kg, about 0.2 mg/kg, about 0.3 mg/kg, about 0.33mg/kg, about 0.5 mg/kg, about 0.75 mg/kg, about 1.0 mg/kg, about 2.0mg/kg, about 3.0 mg/kg, about 4.0 mg/kg, about 5.0 mg/kg, about 6.0mg/kg, about 9.0 mg/kg, or about 15 mg/kg.

In exemplary embodiments, a non-human animal may receive about a 2 mg/kgdosage of grapiprant. In another exemplary embodiment, a non-humananimal may receive about a 3 mg/kg dosage of grapiprant. In oneexemplary embodiment, a non-human animal may receive about a 4 mg/kgdosage of grapiprant. In another exemplary embodiment, a non-humananimal may receive greater than about a 2 mg/kg dosage of grapiprant. Inyet another exemplary embodiment, a non-human animal may receive lessthan about a 50 mg/kg dosage of grapiprant. In one exemplary embodiment,a non-human animal may receive less than about a 6 mg/kg dosage ofgrapiprant.

In one embodiment, a non-human animal may receive about a 3 mg/kg dosageof grapiprant. In one embodiment, a non-human animal may receive about a6 mg/kg dosage of grapiprant. In one embodiment, a non-human animal mayreceive about a 9 mg/kg dosage of grapiprant. In one embodiment, anon-human animal may receive about a 15 mg/kg dosage of grapiprant. Inone embodiment, a non-human animal may receive about a 50 mg/kg dosageof grapiprant.

The dosing may be divided into multiple treatment regimens, depending onseverity of the indications of the non-human animal. For example, insome embodiments, the pharmaceutical composition may be administered tothe non-human animal in need thereof at least once daily, such as atleast twice daily, at least thrice daily, or at multiple times each day.In other embodiments, the pharmaceutical composition may be administeredto the non-human animal in need thereof about 10 hours to about 18 hoursbefore surgery, such as about 11 hours, about 12 hours, about 13 hours,about 14 hours, about 15 hours, about 16 hours, about 17 hours, or about18 hours before surgery.

The dosage of grapiprant may also be expressed in terms of a dosagerate; that is, the total amount of grapiprant provided to a non-humananimal per kilogram bodyweight over the course of a 24-hour period(mg/kg/day). In some embodiments, the grapiprant dosage rate may rangefrom about 1.5 mg/kg/day to about 12 mg/kg/day. In other embodiments,the grapiprant dosage may be at least about 0.2 mg/kg/day, about 0.4mg/kg/day, about 0.6 mg/kg/day, about 0.66 mg/kg/day, about 1.0mg/kg/day, about 1.5 mg/kg/day, about 2.0 mg/kg/day, about 4.0mg/kg/day, about 6.0 mg/kg/day, about 8.0 mg/kg/day, about 10.0mg/kg/day, about 12.0 mg/kg/day, or about 15.0 mg/kg/day. In exemplaryembodiments, the non-human animal may receive grapiprant at a dosagerate of about 1 to about 8 mg/kg/day. In other exemplary embodiments,the non-human animal may receive grapiprant at a dosage rate of about 2to about 4 mg/kg/day. In one embodiment, the non-human animal mayreceive grapiprant at a dosage rate of about 3 to about 15 mg/kg/day.

Dosing may be in the form of solid or liquid formulations. For example,some non-human animals, such as dogs, may receive one or more solid oralformulations, such as a pharmaceutical composition formulated foradministration via capsules, gel caps, gel-like liquids (i.e., viscousliquids), solutions, suspensions, pills, caplets, tablets, or othersolid, liquid, or nebulized forms. For example, the capsules or otherforms may include different concentrations of grapiprant to enabledosing of non-human animals of a plurality of bodyweights. By way ofexample only, capsules may be manufactured with a grapiprantconcentration of 20 mg per capsule, 35 mg per capsule, or 75 mg percapsule. As a result, different combinations of capsules may beadministered to the non-human animals in need of treatment. By way ofexample only, a non-human animal weighing about 15 kg and placed on a3-mg/kg treatment regimen would use about 45 mg of grapiprant per dose.Accordingly, the non-human animal may receive two 20-mg capsules toprovide a dose of grapiprant that is close to 45 mg (i.e., within 5 and10 milligrams of the desired dose based on bodyweight or within a dosingband). Non-human animals of other sizes placed on other treatmentregimens may be similarly treated to provide an efficacious amount ofgrapiprant.

The administering may achieve a C_(max) of grapiprant of about 375 ng/mLto about 10000 ng/mL at a T_(max) of about 0.4 to about 3.4 hours, suchas a C_(max) of grapiprant of 750 about ng/mL to about 8000 ng/mL, or ofabout 1300 ng/mL to about 4000 ng/mL. In other embodiments, the C_(max)of grapiprant may also be achieved at a T_(max) of about 0.7 to about1.7 hours, such as a T_(max) of about 0.5 to about 1.0 hours.

In some embodiments, the C_(max) may vary in different non-humananimals, therefore the C_(max) could be even higher. For example, theC_(max) may be about 375 ng/mL, about 400 ng/mL, about 500 ng/mL, about600 ng/mL, about 700 ng/mL, about 800 ng/mL, about 900 ng/mL, about 1000ng/mL, about 1500 ng/mL, about 2000 ng/mL, about 2500 ng/mL, about 3000ng/mL, about 3500 ng/mL, about 4000 ng/mL, about 4500 ng/mL, about 5000ng/mL, about 5500 ng/mL, about 6000 ng/mL, about 6500 ng/mL, about 7000ng/mL, about 7500 ng/mL, about 8000 ng/mL, about 8500 ng/mL, about 9000ng/mL, about 9500 ng/mL, or about 10000 ng/mL. In exemplary embodiments,the C_(max) of grapiprant may be 375 ng/mL to 10000 ng/mL. In otherexemplary embodiments, the C_(max) of grapiprant may be 750 ng/mL to8000 ng/mL. In yet other exemplary embodiments, the C_(max) ofgrapiprant may be 1300 ng/mL to 4000 ng/mL. In some embodiments, theC_(max) of grapiprant may be greater than 375 ng/mL. In otherembodiments, the C_(max) of grapiprant may be less than 10000 ng/mL.

In other embodiments, the T_(max) may occur in an individual non-humananimal at a 30-minute, 1-hour, or 2-hour time interval, with the rangebeing from about 30 minutes to 2 hours to reach T_(max). The T_(max) maybe about 0.4 hours, about 0.5 hours, about 0.6 hours, about 0.7 hours,about 0.8 hours, about 0.9 hours, about 1.0 hours, about 1.1 hours,about 1.2 hours, about 1.3 hours, about 1.4 hours, about 1.5 hours,about 1.6 hours, about 1.7 hours, about 1.8 hours, about 1.9 hours,about 2.0 hours, about 2.2 hours, about 2.4 hours, about 2.6 hours,about 2.8 hours, about 3.0 hours, about 3.2 hours, or about 3.4 hours.In one embodiment, the T_(max) may be from 0.4 to 3.4 hours. Inexemplary embodiments, the T_(max) may be from 0.7 to 1.7 hours. Inother exemplary embodiments, the T_(max) may be from 0.5 to 1.0 hours.In some embodiments, the T_(max) may be greater than 0.4 hours. In otherembodiments, the T_(max) may be less than 3.4 hours.

The AUC may range from about 1000 hr*ng/mL to about 11000 hr*ng/mL. Inexemplary embodiments, the AUC may range from about 1000 hr*ng/mL toabout 1500 hr*ng/mL, from about 1500 hr*ng/mL to about 2000 hr*ng/mL,from about 2000 hr*ng/mL to about 2500 hr*ng/mL, from about 2500hr*ng/mL to about 3000 hr*ng/mL, from about 3000 hr*ng/mL to about 3500hr*ng/mL, from about 3500 hr*ng/mL to about 4000 hr*ng/mL, from about4000 hr*ng/mL to about 4500 hr*ng/mL, from about 4500 hr*ng/mL to about5000 hr*ng/mL, from about 5000 hr*ng/mL to about 5500 hr*ng/mL, fromabout 5500 hr*ng/mL to about 6000 hr*ng/mL, from about 6000 hr*ng/mL toabout 6500 hr*ng/mL, from about 6500 hr*ng/mL to about 7000 hr*ng/mL,from about 7000 hr*ng/mL to about 7500 hr*ng/mL, from about 7500hr*ng/mL to about 8000 hr*ng/mL, from about 8000 hr*ng/mL to about 8500hr*ng/mL, from about 8500 hr*ng/mL to about 9000 hr*ng/mL, from about9000 hr*ng/mL to about 9500 hr*ng/mL, from about 9500 hr*ng/mL to about10000 hr*ng/mL, from about 10000 hr*ng/mL to about 10500 hr*ng/mL, orfrom about 10500 hr*ng/mL to about 11000 hr*ng/mL.

The half-life (t_(1/2)) may range from about 1.5 hours to about 9 hours.In some embodiments, the t_(1/2) may range from about 3 hours to about14 hours. In exemplary embodiments, the t_(1/2) may range from about 1.5hours to about 2 hours, from about 2 hours to about 2.5 hours, fromabout 2.5 hours to about 3 hours, from about 3 hours to about 3.5 hours,from about 3.5 hours to about 4 hours, from about 4 hours to about 4.5hours, from about 4.5 hours to about 5 hours, from about 5 hours toabout 5.5 hours, from about 5.5 hours to about 6 hours, from about 6hours to about 6.5 hours, from about 6.5 hours to about 7 hours, fromabout 7 hours to about 7.5 hours, from about 7.5 hours to about 8 hours,from about 8 hours to about 8.5 hours, from about 8.5 hours to about 9hours, from about 9 hours to about 9.5 hours, from about 9.5 hours toabout 10 hours, from about 10 hours to about 10.5 hours, from about 10.5hours to about 11 hours, from about 11 hours to about 11.5 hours, fromabout 11.5 hours to about 12 hours, from about 12 hours to about 12.5hours, from about 12.5 hours to about 13 hours, from about 13 hours toabout 13.5 hours, or from about 13.5 hours to about 14 hours.

The duration of administration can and will vary. In general, thepharmaceutical composition may be administered to a non-human animal inneed thereof for about 6 days to about 9 months. In particularembodiments, the pharmaceutical composition may be administered to anon-human animal in need thereof for about 6 days, for about 7 days, forabout 8 days, for about 9 days, for about 10 days, for about 11 days,for about 12 days, for about 13 days, for about 14 days, for about 15days, for about 16 days, for about 17 days, for about 18 days, for about19 days, for about 20 days, for about 21 days, for about 22 days, forabout 23 days, for about 24 days, for about 25 days, for about 26 days,for about 27 days, or for about 28 days.

In exemplary embodiments, the pharmaceutical composition may beadministered to a non-human animal in need thereof for about 9 days toabout 21 days. In other exemplary embodiments, the pharmaceuticalcomposition may be administered to a non-human animal in need thereoffor about 12 days to about 14 days. In some embodiments, thepharmaceutical composition may be administered to a non-human animal inneed thereof for about 1 month, about 2 months, about 3 months, about 4months, about 5 months, about 6 months, about 7 months, about 8 months,or about 9 months. In one embodiment, the pharmaceutical composition maybe administered to a non-human animal in need thereof for about 28 days.

In some exemplary embodiments, the pharmaceutical formulation mayadministered twice daily at a dosage of about 2 mg/kg/day to about 4mg/kg/day for about 9 days to about 21 days, and the administering mayachieve a C_(max) of grapiprant of about 750 ng/mL to about 2200 ng/mLat a T_(max) of about 0.7 to about 1.7 hours.

In some embodiments, the pharmaceutical composition may comprise aliquid oral formulation that may be used in a manner similar to theabove solid oral formulation. Additionally, the liquid formulations maybe administered in a syringe or sprayed on to the animal's food, treats,or chews. Where the non-human animal is livestock, the pharmaceuticalcomposition may be sprayed onto or incorporated into the feed. Forexample, the liquid formulation may be prepared to comprise about 20mg/mL, about 30 mg/mL, about 40 mg/mL, or about 60 mg/mL of grapiprantwithin the liquid formulation.

Similar to the solid formulations discussed above, the differentconcentrations of the liquid formulation may be used to dosing non-humananimals of a plurality of bodyweights. As a result, different volumes ofthe different solutions may be administered to the non-human animals toprovide a dose of grapiprant. By way of example only, a non-human animalweighing about 15 kg and placed on a 3 mg/kg treatment regimen would useabout 45 mg of grapiprant per dose. Accordingly, the non-human animalmay receive about 2.3 mL of the 20 mg/mL solution or 1.1 mL of the 40mg/mL solution to provide a dose of grapiprant close to 45 mg.Similarly, if the same non-human animal was placed on a 4.5 mg/kgtreatment regimen, the animal could receive 2.3 mL of the 30 mg/mLsolution or 1.1 mL of the 60 mg/mL solution to provide a dose ofgrapiprant close to 67.5 mg (i.e., the dose a 15 kg animal shouldreceive on this treatment regimen). Other non-human animals of othersizes placed on other treatment regimens may be similarly treated toprovide an efficacious amount of grapiprant.

In some embodiments, the pharmaceutical composition may be administeredusing any one of a plurality of routes of administration. Thepharmaceutical composition may be orally, parenterally, and/or topicallyadministered. For example, the pharmaceutical composition may be orallyformulated in a liquid and/or a solid formulation so that thecomposition may be administered using at least one of a spray, asyringe, a pill, a tablet, a caplet, a gel-cap, or an otherwiseliquid-based administration scheme.

In other embodiments, the pharmaceutical composition may be formulatedfor administration via subcutaneous, intradermal, intravenous,intramuscular, intracranial, intraperitoneal, or intrathecaladministration (e.g., via an injection or composition-dispensing pump).The pharmaceutical composition may be formulated as a parenterallyadministered depot formulation that can be configured for extendedrelease of the grapiprant (e.g., release over the period of multipledays to multiple months). Moreover, the pharmaceutical composition maybe administered as a gel that contacts the skin or other tissue of theanimals and is accordingly absorbed therethrough. Alternatively, thepharmaceutical composition may be administered using an electrophoreticsystem to drive the composition into circulation of the non-humananimal. In yet other embodiments, the pharmaceutical composition may beformulated for transdermal and/or transmucosal administration (e.g., viaa buccal film or patch that is applied to the inner cheek of thenon-human animal). In addition, in some embodiments, the pharmaceuticalcomposition may be administered intranasally or in the form of one ormore suppositories. In some embodiments, the pharmaceutical compositionmay be formulated as an implant that may be disposed within the softtissue of the non-human animals. For example, the composition-containingimplant may be implanted into the cutaneous, subcutaneous, and/or muscletissue of the non-human animal for extended release. Moreover, thepharmaceutical composition may also be formulated to be administered tothe skin of the non-human animal in a “spot-on” manner. In yet otherembodiments, the pharmaceutical composition may be formulated for anyother suitable route of administration known in the art. In exemplaryembodiments, the pharmaceutical formulation may be administered orallyand may be selected from any pharmaceutical formulation described abovein Section (I).

In some embodiments, the pharmaceutical composition may be administeredto the non-human animal as a part of a daily feeding regimen. Forexample, the pharmaceutical composition may be formulated to be mixedwith the feed or other food product intended for the non-human animalsuch that, as the non-human animal intakes its daily food (e.g., kibbleor soft food), the non-human animal also consumes the pharmaceuticalcomposition. In particular, the pharmaceutical composition may beformulated as a liquid or a powder so that before feeding the non-humananimal, the pharmaceutical composition may be applied (e.g., sprayed)onto the food. Moreover, in some embodiments, the food provided to thenon-human animals may be provided with the pharmaceutical compositionalready added such that the non-human animal's caretaker need onlyprovide the medicated food to the non-human animal.

Other food products provided to the non-human animal may be supplementedwith the grapiprant composition. For example, soft or hard treats orchews (e.g., rawhide or other animal-based products given to non-humananimals for enjoyment and/or enrichment) may be supplemented with thegrapiprant composition, where the grapiprant composition may either beincorporated into the treat or chew or sprayed onto the treat or chew.In some aspects, the treats or chews may be purchased in a form thatalready includes the grapiprant composition. In other aspects, thegrapiprant composition may be later added to the treats or chews by theindividual feeding the animal.

Moreover, in some embodiments, the kibble, treats, and/or chews may bemixed with a maintenance level dosage of the grapiprant-containingcomposition. Preferably, an animal receiving the maintenance leveldosage is able to maintain a certain level of food consumption. Forexample, as discussed above, a maintenance dose (e.g., 0.2 mg/kg) can beprovided to the animals on a regular or irregular basis to provide lowerdoses of the active ingredient. By providing these maintenance doseswith the food products (e.g., kibble), treats, and/or chews, the animalscan relatively enjoy the experience of receiving the maintenance dosessuch that little to no active ingredient is lost in the administrationprocess.

Definitions

The compounds described herein have asymmetric centers. Compounds of thepresent disclosure containing an asymmetrically substituted atom may beisolated in optically active or racemic form. All chiral,diastereomeric, racemic forms and all geometric isomeric forms of astructure are intended, unless the specific stereochemistry or isomericform is specifically indicated.

The term “animal” designates non-human animals, such as “livestock” and“companion animals.” “Livestock” includes cows, sheep, pigs, poultry(e.g., chickens, turkeys, quail, etc.) goats, llamas, and other similaranimals. The term “companion animal” includes, but is not limited, todogs, cats, rabbits, ferrets, horses, and hamsters. In exemplaryembodiments, the companion animal is a dog, cat, or horse.

The term “composition” applies to any solid object, semisolid, or liquidcomposition designed to contain a specific pre-determined amount (dose)of a certain ingredient, for example, an active pharmaceuticalingredient, as previously mentioned and as discussed below. Suitablecompositions may be pharmaceutical drug delivery systems, includingthose for oral administration, buccal administration, rectaladministration, topical or mucosal administration, or subcutaneousimplants, or other implanted drug delivery systems; or compositions fordelivery minerals, vitamins and other nutraceuticals, oral care agents,flavorants, flavor-masking agents, and the like. In one embodiment, thecompositions are generally liquid, however they may contain solid orsemi-solid components. Generally, the dosage form is an orallyadministered system for delivering a pharmaceutical active ingredient tothe alimentary canal of a companion animal.

The phrase “therapeutically effective amount” refers to an amounteffective, at dosages and for periods of time necessary, to achieve thedesired therapeutic result. A therapeutically effective amount ofgrapiprant may be determined by a person skilled in the art (e.g., aveterinarian) and may vary according to factors such as the clinicalstate, age, sex, and weight of the companion animal, bioavailability ofgrapiprant, and the ability of the active agent(s) to elicit a desiredresponse in the companion animal. A therapeutically effective amount isalso one in which any toxic or detrimental effects of the activeagent(s), are outweighed by the therapeutically beneficial effects. Atherapeutically effective amount also encompasses an amount that iseffective, at dosages and for periods of time necessary, to achieve thedesired result (e.g., weight gain through the addition of lean musclemass).

The term “q.s.” means to add a quantity (e.g., volume or mass) of aningredient until the final amount (e.g., volume or mass) is reached.

The term “w/v” designates a concentration of a substance as measured inweight of the substance per volume of a solution or composition.

When introducing elements of the present disclosure or theembodiments(s) thereof, the articles “a,” “an,” “the,” and “said” areintended to mean that there are one or more of the elements. The terms“comprising,” “including,” and “having” are intended to be inclusive andmean that there may be additional elements other than the listedelements.

Having described the disclosure in detail, it will be apparent thatmodifications and variations are possible without departing from thescope of the disclosure defined in the appended claims.

EXAMPLES

The following examples are included to demonstrate certain embodimentsof the disclosure. It should be appreciated by those of skill in the artthat the techniques disclosed in the examples represent techniquesdiscovered by the inventors to function well in the practice of thedisclosure. Those of skill in the art should, however, in light of thepresent disclosure, appreciate that many changes can be made in thespecific embodiments that are disclosed and still obtain a like orsimilar result without departing from the spirit and scope of thedisclosure, therefore all matter set forth is to be interpreted asillustrative and not in a limiting sense.

Example 1—Palatability Study of Grapiprant in Dogs

Fifty privately-owned pet dogs were used in the study. The dogs were amixture of breeds, sexes (male and female, all neutered), aged from 1.5years to 13 years, and weighing from 14.5 to 113 pounds. The treatmentwas a flavored tablet containing 12.5 mg of grapiprant. Four differentflavored tablets (A, B, C, and D) were used for each dog, regardless ofbodyweight. The flavors used were beef, pork liver, and two other flavorcombinations. All test articles were manufactured by ArgentaManufacturing, Manurewa, Auckland, New Zealand. Composition of the testarticles are shown at Table 1.

TABLE 1 Grapiprant formulations used in the palatability study. Formula(% w/w) Material Name Tablet A Tablet B Tablet C Tablet D Grapiprant 2.42.4 2.4 2.4 Microcrystalline 40.0 35.0 35.0 35.0 Cellulose LactoseSupertab 45.6 40.6 40.6 40.6 Spray Dried Sodium Starch 5.0 5.0 5.0 5.0Glycolate Flavorant Protex 3496 “A” Flavor Powder- Flavor Powder- FlavorPowder- Flavor Powder Spray-Dried Artificial Artificial 5.0 Pork LiverBeef PC-0125 PC-0335 15.0 15.0 15.0 Colloidal Silicon 0.5 0.5 0.5 0.5Dioxide Magnesium 1.5 1.5 1.5 1.5 Stearate

TABLE 2 Palatability Evaluation, Results. Tablet A Tablet B Tablet CTablet D Response N = 12 N = 13 N = 12 N = 13 Dog ate completely, 1(8.3%)  11 (84.6%)  8 (66.7%) 7 (53.8%) immediately Dog played with, but2 (16.7%) 1 (7.7%)  2 (16.7%) 3 (23.1%) eventually consumed Dog put inmouth, but 3 (25.0%) 0 (0.0%) 1 (8.3%) 1 (7.7%)  spit out Dog completelyrefused 6 (50.0%) 1 (7.7%) 1 (8.3%) 2 (15.4%) Acceptability/Palatability3 (25.0%) 12 (92.3%) 10 (83.3%) 10 (76.9% ) Palatable to dog, as 3(25.0%) 11 (84.6%) 10 (83.3%) 9 (69.2%) assessed by owner Unpalatable todog, as 9 (75.0%) 2 (15.4%)  2 (16.7%) 4 (30.8%) assessed by owner

This study was a randomized, blinded, non-GCP clinical trial. Based onthese data, tablet B was preferred by the test animals, followed bytablets C and D. Tablet A was rejected by most (75.0%) of dogs tested.In treatment groups A and C, the owner's assessment of palatabilitycoincided with whether the dogs consumed the tablet. In these two cases,the dogs did eventually consume the tablet; however, the owners reportedthat their dogs probably did not find the tablets palatable. Overall,Tablet B was found to be the most palatable by both metrics. Nostatistical tests were applied to these data.

Example 2—Another Palatability Study of Grapiprant in Dogs

A total of 40 privately-owned dogs were presented at two veterinaryclinics. The dogs were a mixture of breeds, sexes (male and female, allneutered except two females), aged from 15 months to 13 years, 10months, and weighing from 19.6 to 109 pounds. In total, four protocoldeviations related to selection criteria.

The treatment was a flavored tablet containing 20 mg of grapiprant. Fourdifferent flavored tablets (A27, A29, A31, and A34) were used for alldogs regardless of weight. The flavoring agent used was derived frompork liver. The tablets differed in the amount (5%, 10% or 15%) offlavoring agent included. Two of the formulations (labeled A27 and A34)evaluated contained 15% pork liver flavoring; the difference between thetwo formulations lies in the surfactant component: Formulation A27contained poloxamer, while Formulation A34 contained sodium laurelsulfate. All test articles were manufactured by Argenta Manufacturing(Manurewa, Auckland, New Zealand) and received by Ricerca Laboratories(Concord, Ohio). Composition of the test articles was as shown in Table3.

TABLE 3 Formulations of grapiprant. Formulation Formulation FormulationFormulation Material* A27 A29 A31 A34 Grapiprant 8.0 8.0 8.0 8.0 Lactose36.0 36.0 41.0 35.0 (Super-Tab) Sodium Starch 5.0 5.0 5.0 5.0 GlycolateMicrocrystalline 28.0 38.0 28.0 28.0 Cellulose Colloidal Silicon 0.5 0.50.5 0.5 Dioxide Magnesium 1.5 1.5 1.5 1.5 Stearate Flavor Powder- 15.05.0 10.0 15.0 Pork Liver Copovidone 5.0 5.0 5.0 5.0 (Kollidon VA64)Surfactant 1.0 1.0 1.0 2.0 *Amounts of each component are listed asweight percent of the total formulation.

After eating the tablet (if applicable), the owner's assessment ofwhether or not he or she believed that the dog found the tabletpalatable was recorded. Dogs were observed for 5 minutes forregurgitation/vomiting, or adverse reactions to the administration(e.g., frothing/foaming, pawing at the mouth, other abnormal behaviors).Dogs' acceptance or rejection of the study product was recorded.

The relationship between each adverse event and the test article wasclassified in relationship to the treatment as follows:

-   -   1=Unrelated: Clearly pre-existing or caused by a specific        extraneous event.    -   2=Possibly related: Possible drug association as suggested by        the relationship of adverse event with treatment and external        events.    -   3=Related: Strong suspicion of drug association when type, time        course, and relationship of adverse event to treatment and        external events are considered.

Data Analysis and Results. Upon entry into the database, categoricalsummary statistics were performed to evaluate the palatability of thefour formulations. The results are summarized in Table 4 below.“Acceptability/Palatability” is defined as the percentage of dogsoffered each formulation who consumed the tablet within the 5-minuteperiod, whether they consumed it readily or played with the tabletbefore consuming it.

TABLE 4 Palatability Evaluation Results Formula- Formula- FormulationFormulation tion A27 tion A29 A31 A34 Dogs' Behavior N = 9 N = 11 N = 10N = 10 Dog ate completely 4 3 4 2 and Immediately Dog played with, but 12 1 3 eventually consumed Dog put in mouth, but 1 1 1 1 spit out Dogcompletely 3 5 4 4 refused Acceptability/ 55.6% 45.5% 50.0% 50.0%Palatability Palatable to Dog, as 55.6% 36.4% 40.0% 50.0% assessed byOwner Unpalatable to Dog, 44.4% 63.6% 60.0% 50.0% as assessed by Owner

Based on these data, Formulation A27 was most acceptable to the testanimals, with 55.6% consuming the tablet without encouragement.Formulations A31 and A34 were accepted and rejected by an equal number(50%) of dogs tested. Formulation A29 was rejected by more than half(55.4%) of dogs tested. Although the palatability estimates did notdiffer appreciably among the four formulations, there may be a trendtoward a dose-response effect in the level of flavoring compound: 5%pork liver powder <[10% pork liver=15% pork liver (sodium laurel sulfateas surfactant)]<15% pork liver (poloxamer). In all but two cases(Formulation A31 and Formulation A29) the owner's assessment ofpalatability coincided with whether or not the dogs consumed the tablet.

In these cases, the dog did eventually consume the tablet; however, theowners reported that the dogs probably did not find the tabletpalatable. No statistical tests were applied to the data. In thissingle-dose study, no adverse events were reported, nor were there anyobservations of unusual behaviors (frothing/foaming, pawing at themouth, other abnormal behaviors).

Example 3—Pharmacokinetic Studies of Grapiprant in Dogs

The objective of this study was to compare the pharmacokinetic profileof several tablet formulations of grapiprant to capsule and aqueousmethylcellulose suspension formulations of grapiprant with 20-mg doses.Appropriate amounts of the test article/excipient mixture were placedinto gelatin capsules to provide each animal with 20 mg of grapiprant.As the grapiprant content in the excipient blend was 45% of the mixtureweight, the total weight of the blend in each capsule was 0.0440 g. Theweight tolerance was within ±0.005 g of the nominal capsule contentweight.

For the aqueous suspensions (Group 2), the vehicle was 0.5% methylcellulose (400 centipoise) in water. A 0.3-mg/mL suspension ofgrapiprant was prepared by adding grapiprant to a calibrated beaker andadding about 60-90% of the 0.5% aqueous methylcellulose to the beaker. Ahomogenizer was used to blend the material. A stir bar was added to thebeaker, and the suspension was stirred for about 5 minutes. The stir barwas removed temporarily to adjust the volume with additional vehicle,and then the suspension was stirred for another 5 minutes before beingtransferred to a labeled dosing container. The formulation wascontinuously stirred at the time of dose administration.

A single oral dose by capsule, suspension, or tablet was given to dogsshown in Table 5.

TABLE 5 Formulations used in this study. Nominal Number of Dose LevelGroup Animals Formulation (mg/kg)* 1 5 20 mg API in capsule 2 2 5 0.3mg/mL methylcellulose suspension 2 Dose volume: 6.67 mL/kg 3 5 20 mgtablet, Formulation A29 2 4 5 20 mg tablet, Formulation A33 2 5 5 20 mgtablet, Formulation A31 2 6 5 20 mg tablet, Formulation 120314-1 2*based on a 10-kg animal. Only whole tablets were used in Groups 3-6.

Statistical analyses were conducted to determine differences in C_(max),T_(max), and AUCmax Values were normalized by mg/kg, and group means andstandard deviations were calculated for numeric data. The actualadministered doses ranged from 1.81 to 2.82 mg/kg, and averaged 2.2mg/kg. No animal was found dead or deemed moribund during the study. Allanimals were normal at the scheduled observation time of 2 hourspostdose. The mean pharmacokinetic parameters, in addition to thedose-normalized C_(max) and AUC_(0-∞), are shown in Table 6.

TABLE 6 Mean Grapiprant Pharmacokinetic Parameters in Male Dogs on Day0. Dose C_(max) AUC_(last) AUC_(0−∞) C_(max)/Dose AUC_(0−∞)/Dose (mg/(ng/ T_(max) (hr*ng/ (hr*ng/ t_(1/2) (ng/mL)/ (hr*ng/ Group kg) mL) (hr)mL) mL) (hr) (mg/kg) mL)/(mg/kg) 1 2.16 804 1.08 1580 1640 5.5 359 724 22.00 751 0.786 1650 1870 4.1 376 937 3 2.2 1200 0.891 2330 2340 2.17 5461070 4 2.2 887 0.995 2050 2320 8.58 409 1160 5 2.29 697 1.4 1690 18605.87 311 757 6 2.14 816 1.09 2020 2560 7.52 377 1150

Following a single oral dose of 20 mg grapiprant capsule to Group 1dogs, the mean T_(max) was 1.08 hours, the mean terminal half-life was5.5 hours, the mean C_(max) was 804 ng/mL, and the mean AUC_(0-∞) was1640 hr*ng/mL. Following a single oral dose of 2 mg/kg grapiprantsuspension to Group 2 dogs, the mean T_(max) was 0.786 hours, the meanterminal half-life was 4.1 hours, the mean C_(max) was 751 ng/mL, andthe mean AUC_(0-∞) was 1870 hr*ng/mL.

After a single oral dose of 20 mg grapiprant tablet, Formulation A29, toGroup 3 dogs, the mean T_(max) was 0.891 hours, the mean terminalhalf-life was 2.17 hours, the mean C_(max) was 1200 ng/mL, and the meanAUC_(0-∞) was 2340 hr*ng/mL. After a single oral dose of 20 mggrapiprant tablet, Formulation A33, to Group 4 dogs, the mean T_(max)was 0.995 hours, the mean terminal half-life was 8.58 hours, the meanC_(max) was 887 ng/mL, and the mean AUC_(0-∞) was 2320 hr*ng/mL. After asingle oral dose of 20 mg grapiprant tablet, Formulation A31, to Group 5dogs, the mean T_(max) was 1.40 hours, the mean terminal half-life was5.87 hours, the mean C_(max) was 697 ng/mL, and the mean AUC_(0-∞) was1860 hr*ng/mL. After a single oral dose of 20 mg grapiprant tablet,Batch 120322-1, to Group 6 dogs, the mean T_(max) was 1.09 hours, themean terminal half-life was 7.52 hours, the mean C_(max) was 816 ng/mL,and the mean AUC_(0-∞) was 2560 hr*ng/mL.

Mean T_(max) was the shortest after dogs were administered grapiprantsuspension, and AUC_(0-∞) was lowest following a grapiprant capsule.Mean t_(1/2) was the shortest, but C_(max) was highest after a tablet(Formulation A29). Mean t_(1/2) was the longest after a tablet(Formulation A33). Mean T_(max) was the longest, but C_(max) was lowestafter a tablet (Formulation A31).

When normalized for dose received, the C_(max) and AUC_(0-∞) weresimilar among groups with only slightly lower absorption (C_(max) andAUC_(0-∞)) in the capsule (Group 1), suspension (Group 2) and the Group5 tablet formulations; however, analysis of variance (ANOVA) indicatedthat there was no significant difference among the groups for C_(max),T_(max) and AUC_(0-∞). FIGS. 1-6 depict the individual serumconcentrations of grapiprant over time for Group 1-6 dogs, respectively.FIG. 7 depicts the combined individual serum concentrations forgrapiprant over time for Groups 1-6 dogs.

After a single nominal 2-mg/kg oral dose of grapiprant using either acapsule, methylcellulose suspension, or one of four different formulatedtablets, pharmacokinetic parameters varied with dose form. Whennormalized for dose received, the C_(max) and AUC_(0-∞) were similaramong groups with only slightly lower absorption (C_(max) and AUC_(0-∞))in the capsule (Group 1), suspension (Group 2) and the Group 5 tabletformulations; however, ANOVA indicated no significant difference amongthe groups for C_(max), T_(max) and AUC_(0-∞).

Example 4—Further Pharmacokinetic Studies of Grapiprant in Dogs

The objective of this study is to determine the pharmacokinetic profileof several tablet formulations of grapiprant administered to male andfemale beagle dogs. The test article was tested in four formulations:Group 1 (Formulation A34), Group 2 (Formulation A27), Group 3(Formulation A29), and Group 4 (Formulation A31), as described above atTable 4 of Example 2. Each tablet formulation of the test articlecontained 20 mg of the active pharmaceutical ingredient in a distinctexcipient mixture. The test article was used without correction forpurity or salt content. A single oral dose with one whole tablet wasgiven to dogs as shown below in Table 7.

TABLE 7 Formulations used in this study. Nominal Number of Dose LevelGroup Animals Formulation (mg/kg)* 1 4/4 20 mg tablet, Formulation A34 22 4/4 20 mg tablet, Formulation A27 2 3 4/4 20 mg tablet, FormulationA29 2 4 4/4 20 mg tablet, Formulation A31 2 *Based on animal bodyweightof 10 kg.

Animals received a detailed clinical evaluation beforeinclusion/randomization on Day −6. Animal bodyweights were within normallimits for beagles of this age and gender. After dosing, no signs ofregurgitation or vomit were observed. Dogs were dosed with a nominal2-mg/kg oral dose of grapiprant using four different formulated tablets.

The actual dose levels for Group 1 ranged from 2.05 to 3.23 mg/kg, Group2 ranged from 1.82 to 3.15 mg/kg, Group 3 ranged from 1.69 to 3.23mg/kg, and Group 4 ranged from 2.05 to 3.15 mg/kg. After a single oraldose of 20-mg grapiprant tablet Formulation A34 to Group 1 dogs, themean T_(max) was 0.975 to 0.992 hour. The mean terminal half-life was5.71 to 6.88 hours. Mean C_(max) was 1750 to 2180 ng/mL, and meanAUC_(0-∞) was 4420 to 4650 hr*ng/mL. After a single oral dose of 20-mggrapiprant tablet Formulation A27 to Group 2 dogs, the mean T_(max) was0.704 to 0.709 hours. The mean terminal half-life was 6.41 to 9.22hours. Mean C_(max) was 1430 to 2460 ng/mL, and mean AUC_(0-∞) was 3190to 5160 hr*ng/mL. After a single oral dose of 20-mg grapiprant tabletFormulation A29 to Group 3 dogs, the mean T_(max) was 0.704 to 0.933hours. The mean terminal half-life was 3.11 to 6.83 hours. Mean C_(max)was 1270 to 1900 ng/mL, and mean AUC_(0-∞) was 2840 to 3530 hr*ng/mL.After a single oral dose of 20 mg grapiprant tablet Formulation A31 toGroup 4 dogs, the mean T_(max) was 0.817 to 0.933 hour. The meanterminal half-life was 5.15 to 7.42 hours. Mean C_(max) was 1430 to 2160ng/mL, and mean AUC_(0-∞) was 2860 to 3410 hr*ng/mL.

FIGS. 8 and 9 depict individual serum concentrations of grapiprant overtime for Group 1 male and female dogs, respectively, tested withFormulation A34. FIGS. 10 and 11 depict individual serum concentrationsof grapiprant over time for Group 2 male and female dogs, respectively,tested with Formulation A27. FIGS. 12 and 13 depict individual serumconcentrations of grapiprant over time for Group 3 male and female dogs,respectively, tested with Formulation A29. FIGS. 14 and 15 depictindividual serum concentrations of grapiprant over time for Group 4 maleand female dogs, respectively, tested with Formulation A31.

FIG. 16 depicts the mean serum concentrations of grapiprant over timefor Group 1 dogs grouped by male and female, as individually shown atFIGS. 8 and 9. FIG. 17 depicts the mean serum concentrations ofgrapiprant over time for Group 2 dogs grouped by male and female, asindividually shown at FIGS. 10 and 11. FIG. 18 depicts the mean serumconcentrations of grapiprant over time for Group 3 dogs grouped by maleand female, as individually shown at FIGS. 12 and 13. FIG. 19 depictsthe mean serum concentrations of grapiprant over time for Group 4 dogsgrouped by male and female, as described individually shown at FIGS. 14and 15. FIG. 20 depicts the mean serum concentrations of grapiprant overtime for Groups 1-4 dogs, as shown at FIGS. 8-19.

No sex difference was observed for any tablet formulation in the study.Mean T_(max) was the longest and variability for AUC was the lowestafter a single oral dose of 20 mg grapiprant tablet Formulation A34.Mean T_(max) was the shortest and t_(1/2) was the longest andvariability for C_(max) was the lowest after a tablet Formulation A27.Tablet Formulation A29 had the lowest mean C_(max) and t_(1/2), but hadthe highest variability for C_(max) and AUC values. Overall, all fourformulations were very similar.

Following a single nominal 2-mg/kg oral dose of grapiprant using fourdifferent formulated tablets, C_(max) among formulations did not differsignificantly (p>0.05), but T_(max) was significantly different(p<0.05). T_(max) of tablet Formulation A34 was significantly longerthan that of tablet Formulations A27, A29, and A31 (p<0.05), no otherpair comparisons differed significantly (p>0.05). AUC_(last) of anytablet formulation did not differ significantly from others when paircomparisons were done (p>0.05).

Example 5—Efficacy Study of Grapiprant in Cats

The objective of this pilot study was to validate that this model/painscoring system can differentiate positive from negative control animalsin controlling the pain and inflammation of onychectomy and to test theeffectiveness of a single dose of grapiprant in controlling the pain andinflammation associated with onychectomy in cats.

Following an acclimation period of 7, 8, or 9 days, a total of 30 adultcats were randomly divided between three treatment groups with tenanimals assigned to each group. All animals were randomly assigned to agiven surgery day such that within each day, an equal number of cats wasfrom each treatment group. Within each day, the order of surgery wasalso randomized. On Day 0, all study animals were administered asubcutaneous injection of butorphanol at a target dose of 0.4 mg/kgbodyweight just prior to treatment administration preceding surgery.

All animals in Group 1 were orally-administered a 6 mg tablet ofPositive Control (Onsior™, robenacoxib), animals in Group 2 wereadministered an empty gelatin capsule as the Negative Control, andanimals in Group 3 were administered a 20 mg tablet of grapiprant(Formulation A34, see Table 4 in Example 2 above) all about 30 minutesbefore onychectomy. A water chaser was not administered followingdosing. On Days 1 and 2, animals in Group 1 were administered a 6 mgtablet or Positive Control, while animals in Groups 2 and 3 wereadministered an empty gelatin capsule. Details of the experimental studydesign are summarized in Table 8.

TABLE 8 Description of Study Design. Number of Group Animals Dose RouteDose 1 10 Positive Control PO Days 0, 1, and 2: (Onsior ™) 6 mg/animal(full 6 mg tablet) 2 10 Negative Control PO Days 0, 1, and 2: (gelatincapsule) 0 mg (empty capsule) 3 10 Grapiprant PO Days 0: ~20 mg/animal(20 mg tablet) (full 20 mg tablet) Formulation A34 Days 1 and 2: 0 mg(empty capsule)

The variables evaluated and the intervals they were determined aresummarized in Table 9.

TABLE 9 Variables Evaluated and Description of Study Design. Study DayActivity/Parameter Throughout General health observations (at least onceper day) Up to Days −9 to −1 Acclimation Day −4 Physical ExaminationsDay −1 Body Weight Measurements Day −1 Randomization Day 0 (0.5 hours, 1hour, and every Sedation Score other hour until animal received twoconsecutive scores of one) Days 0 (0.5, 1, 3, 5, and 8 Analgesia Scoringhours-post extubation), 1 (24 hours post-dose Day 0, 1200 to 1400, and1700 to 1900), 2 (48 hours post-dose Day 0, 1200 to 1400, and 1700 to1900), and 3 (72 hours post-dose Day 0)

On Day 0, cats were orally-dosed with the Positive Control (Group 1),Negative Control (Group 2), or grapiprant (Group 3) about 30 minutesbefore onychectomy. On Days 1 and 2, animals in Group 1 were orallydosed with Positive Control while animals in Groups 2 and 3 were giventhe Negative control.

Surgical Procedure. Surgeries for the study were staggered overdifferent days. Study animals were divided into sequences of 9, 9, and12 cats to allow for surgeries to be performed over three days. The dayof surgery was considered to be Day 0 for that sequence. All animalswere given a subcutaneous injection of butorphanol at a target dose of0.4 mg/kg bodyweight just before treatment administration precedingsurgery. Cats were premedicated with acepromazine (target 0.05 mg/kg,subcutaneous) and anesthesia was induced with propofol (not more than 8mg/kg, intravenous slowly to effect). Anesthesia was maintained byisoflourane (0.5 to 5% in 100% oxygen). Animals were intubated at thediscretion of veterinary staff. During surgical procedures, each animalwas monitored for at least heart rate and respiratory rate. Onychectomyof the front paws on all animals was performed with surgical scalpel andsoft tissues and skin were closed with GLUture™ Topical Tissue Adhesiveat the conclusion of surgery. Following surgery, each animal wasperiodically monitored until full recovery. Each cat was extubated whenthe swallowing reflex was observed to return. Extubation time (timezero=T₀) was recorded as the end of surgery.

All study animals were scored for sedation and analgesia the followingschedule in Table 10.

TABLE 10 Sedation and analgesia scoring schedule. Day Time AssessmentComments 0 0 Extubation 30 ± 10 min Sedation score Performed by maskedAnalgesia score Assessor  1 hour ± 10 min Sedation score As aboveAnalgesia score  3 hour ± 30 min Sedation score (or until animal Asabove received two consecutive scores of 1) Analgesia score  5 hour ± 30min Sedation score (or until animal As above received two consecutivescores of 1)  8 hour ± 30 min Sedation score (or until animal As abovereceived two consecutive scores of 1) 12 hour ± 30 min Analgesia scoreAs above 16 hour ± 30 min Analgesia score As above 20 hour ± 30 minAnalgesia score As above 24 hour ± 30 min Analgesia score As above 1 24hour ± 1 hour Analgesia score As above (post-Day 0) Mid-day Analgesiascore As above 1200-1400 hours Early evening Analgesia score As above1700-1900 hours 2 4 hour ± 1 hour Analgesia score As above (post-Day 0)Mid-day Analgesia score As above 1200-1400 hours Early evening Analgesiascore As above 1700-1900 hours 3 72 hour ± 1 hour Analgesia score Asabove (post-Day 0)

Each study animal was observed and given a sedation score at specifiedtime points during the study or until the animal received twoconsecutive scores of 1. A five-point scoring system was used to scoresedation post-operatively.

Analgesia scores were given at specified time points during the study.The masked assessor observed the cat in its cage and then gentlypalpated the surgical site to assess pain. Analgesia was scored by thesame assessor during the first 8 hours of the study. Observations after8 hours were made by alternative assessors, as necessary.

Results. The cat was evaluated as the experimental unit. Differencesbetween treatment groups were evaluated at alpha of 0.05. Sedations andanalgesia scores were evaluated using methods appropriate for ordinaldata measured repeatedly over time, such as the GLIMMIX procedure (SASInstitute, Cary, N.C., version 9.3). A multinomial distribution wasassumed and a cumulative logit link used. The model included treatmentgroup, time, and the group by time interaction as fixed effects. Giventhe unequal spacing of the time intervals, a compound symmetric (CS) orheterogeneous CS structure was assumed for the covariance matrix,depending on the Akaike's Information Criterion. If the group by timeinteraction was significant, within time comparisons were made,comparing groups in a pairwise fashion. If the interaction was notsignificant, the main effect of group was evaluated. If the main effectof group was significant, pairwise comparisons among groups were made.If the algorithm used in the model fitting process did not converge,alternative models were evaluated. Mean values are used to summarize theresults. Additionally, each cat was categorized as either a success orfailure based on the need for pain intervention. The percent ofsuccesses within treatment groups was evaluated using methodsappropriate for binary outcomes, assuming a binomial distribution andlogit link. The model included treatment group as a fixed effect. If theeffect of treatment was significant, pairwise comparisons between groupswere made. The percent failure within group and 95% confidence intervalsare presented. Intervention results were also evaluated using methodsappropriate for binary data measured repeatedly over time. A binomialdistribution was assumed and a logit link used.

All animals were successfully dosed the specified test formulation onDay 0 with the Positive Control, Negative Control, and grapiprantformulation. Beginning at the one hour time point following the surgicalintervention and continuing for the duration of the observation period(72 hours), animals in the Negative Control group consistently hadhigher analgesia scores than their counterparts in the Positive Controland grapiprant groups. However, this trend only reached statisticalsignificance at the Day 1 mid-day time point (P<0.05). These findingswere also found when three animals with non-treatment related extremityfindings (swollen paws and trauma to paw) were excluded from theanalysis, but the differences did not achieve statistical significanceat any time point. The animals in the placebo group required morefrequent, and a greater total number of “rescue” analgesia (butorphanol)than those in either of the other two groups, although these differenceswere not statistically significant.

In summary, administration of the test article grapiprant was associatedwith decreased analgesia scores (reduced pain), without an apparenteffect on sedation scores when compared to placebo controls. Theserelationships were similar to those found for the Positive Controltreatment.

Example 6—Grapiprant Oral Tablet for Treating Osteoarthritis in Dogs

In a masked, multi-centered dose ranging field study, dogs withnaturally occurring osteoarthritis were treated with grapiprant at 2mg/kg once daily (SID), 5 mg/kg once daily, 4 mg/kg twice daily (BID),or placebo twice daily to assess the control of pain and inflammation.Dogs were randomly assigned to one of the four treatment groups andtreated orally with Formulation A34 tablets of 20, 60, and 100 mg, orplacebo tablets (matched to the 4 mg/kg group). (See above at Example 2,Table 4). The control of pain and inflammation was assessed by the ownerusing the validated Canine Brief Pain Inventory (CBPI) assessment tool,which consists of a pain severity score (PSS), a Pain Interference Score(PIS), and an Owner Impression of dog's quality of life. CBPI scoring,as well as a veterinarian assessment of osteoarthritis, were conductedat Study Day 0 (baseline) and Days 7, 14, 21, and 28.

In total 476 dogs were screened in the study with 108 screen failures,resulting in 336 dogs in the per protocol population. Table 11 lists thenumber of animals from each group in the per protocol population.

TABLE 11 Study population numbers per treatment group. Treatment GroupPer Protocol Population Placebo 83 2 mg/kg SID 86 5 mg/kg SID 82 4 mg/kgBID 85

There was no difference in median PSS or PIS scores among the groups atbaseline. A positive response (decreases in the PSS and PIS scores) wasobserved in all treatment groups with statistically significantdifferences seen in the 2 and 5 mg/kg SID groups compared to placebo. OnDays 14, 21, and 28, the PSS scores differed significantly (p≤0.05)among groups, with statistically significant greater improvements forthe 2 mg/kg SID group in pairwise comparisons to placebo on Days 14 and28, and on Days 14, 21, and 28 for the 5 mg/kg group. For the PISscores, the groups differed significantly (p≤0.05) on Day 28 withstatistically significant greater improvements for the 2 and 5 mg/kg SIDgroups in pairwise comparisons to placebo. The descriptive statisticsfor the PSS and PIS scores from the CBPI are given in Tables 12 and 13.

The dose groups had similar efficacy with no benefit from increasing thedose, as shown in the descriptive statistics from the PSS and PIS scoresfrom the CBPI (Tables 12 and 13). The median decreases from baseline PSSscores were 48, 48.53, and 44.44 for the 2 mg/kg SID, the 5 mg/kg SID,and the 4 mg/kg BID dose groups, respectively, compared to only 28 forthe placebo group. For the PIS scores, the median decreases frombaseline were 53.25, 55.27, and 52.27 for the 2 mg/kg SID, the 5 mg/kgSID, and the 4 mg/kg BID dose groups, respectively, compared to only39.13 for the placebo group. These data suggest that the 2 and 5 mg/kgSID doses were both effective and that twice daily dosing added nobenefit.

TABLE 12 Pain Severity Scores (PSS) descriptive statistics. 2 mg/ 4 mg/5 mg/ p- Study Day Placebo kg SID kg BID kg SID value* Day 0 N 83 86 8582 0.5217 Median 5.00 5.25 5.25 5.00 (r) p-value+ 0.9902 0.9999 0.5325 %N 79 84 83 81 Change*: Median −14.29 −24.26 −17.39 −22.22 0.1024 Day 7p-value+ 0.0719 0.9452 0.2785 (r) % N 81 85 80 81 0.0441 Change*: Median−22.22 −32.00 −28.29 −30.00 (r) Day 14 p-value+ 0.0380 0.1739 0.0370 % N75 81 77 77 0.0478 Change*: Median −25.93 −35.00 −38.89 −44.44 (r) Day21 p-value+ 0.1260 0.1102 0.0207 % N 77 84 75 78 0.0109 Change*: Median−28.00 −48.00 −44.44 −48.53 (r) Day 28 p-value+ 0.0250 0.1028 0.0048*Overall p-values generated by analysis of variance with terms fortreatment, site and treatment by site interaction; +Pairwise comparisonsto Placebo; **Percent Change from Day 0

TABLE 13 Pain Interference Score (PIS) descriptive statistics 2 mg/kg 4mg/kg 5 mg/kg p- Study Day Placebo SID BID SID value* Day 0 N 83 86 8582 0.5434 Median 5.67 6.00 5.83 5.50 (r) p-value+ 0.7963 0.9902 0.3876 %N 79 84 83 81 Change**: Median −15.91 −27.97 −28.57 −26.92 0.3295 Day 7p-value+ 0.3039 0.4652 0.1953 (r) % N 81 85 80 81 0.2033 Change**:Median −27.45 −33.33 −35.24 −40.00 (r) Day 14 p-value+ 0.2817 0.36930.0836 % N 75 81 77 77 0.1166 Change**: Median −35.71 −47.83 −42.11−53.85 (r) Day 21 p-value+ 0.1777 0.2755 0.0541 % N 77 84 75 78 0.0321Change**: Median −39.13 −53.25 −52.27 −55.27 (r) Day 28 p-value+ 0.04160.1146 0.0195 *Overall p-values generated by analysis of variance withterms for treatment, site and treatment by site interaction; +Pairwisecomparisons to Placebo; **Percent Change from Day 0

Based on the results of the PSS and PIS scores of the CBPI, the 2 mg/kgSID and the 5 mg/kg SID dose are equally effective in controlling thepain and inflammation of osteoarthritis in dogs. The preliminaryestimated mean effective canine dose of 2 mg/kg SID based on preliminaryextrapolation calculations from pharmacokinetic/pharmacodynamic studiesin human Phase 1 and 2 trials, protein binding, and comparative receptoraffinity of grapiprant between humans and dogs, was confirmed in thedose ranging field study. The dose was effective in field conditions ofuse without food restrictions. Thus, these results support the use of 2mg/kg SID given with or without food for the control of pain andinflammation associated with osteoarthritis in dogs.

Example 7-9—Month Oral Toxicity Study of Grapiprant in Dogs with 4-WeekRecovery (06NG032)

Grapiprant was administered orally, once daily, for nine consecutivemonths to Beagle dogs at doses of 0 (0.5% methylcellulose), 1, 6, and 50mg/kg/day in a dose volume of 5 mL/kg. Four animals per sex were used ineach dose group and two additional animals per sex in the 50 mg/kg dosegroup for recovery purposes. Clinical signs and food consumption wereassessed daily. Bodyweight was recorded weekly. Ophthalmologicexamination was performed on Weeks 20 and 38 of the dosing phase andWeek 4 of the recovery phase. Electrocardiograms were recorded on Weeks13, 26, and 38 of the dosing phase and Week 4 of the recovery phase.Hematology, coagulation, and serum chemistry parameters were monitoredon Weeks 13, 26, and 39 of the dosing phase and Week 4 or 5 of therecovery phase. Urinalyses were performed on Week 37 of the dosing phaseand Week 3 or 4 of the recovery phase. Serum drug concentrations ofgrapiprant were measured at 0.5, 1, 2, 4, 8, and 24 hours post-dose onDay 1 (50 mg/kg only) and at Week 38. At the end of the dosing orrecovery period, dogs were euthanized and necropsied. After grossexamination, selected organs were weighed, and a comprehensive set oftissues was collected and processed for microscopic examination.

There were no deaths or drug related effects on bodyweight, foodconsumption, ophthalmology, electrocardiograms, hematology, coagulation,organ weights, or gross pathological findings up to 50 mg/kg during the9-month dosing period. Gastrointestinal effects such as loose or mucousstool, which sometimes included slight bloody or red material wereobserved in all groups including the control. The incidence was higherin some animals at 1-50 mg/kg compared with that in the control animals.A significant decrease in mean serum albumin was observed at Weeks 26and 39 (up to −14% vs. control value) at 50 mg/kg and in meanalbumin/globulin (A/G) ratio at Week 39 at 6 mg/kg (−16%). Individually,there was a dose-related trend for increase in incidence and decrease inonset time, with decreases in albumin (up to −41% vs. pre-study), totalprotein (up to −30%) and/or calcium (up to −18%). These findingsreturned to normal range after a one-month reversal. The serum parameterchanges were recovered at the end of the recovery period. There were nonoteworthy findings during or at the end of the 4-week recovery period.

After single or repeated oral administrations, there were no sex relateddifferences in the systemic exposure to grapiprant, no accumulation ofgrapiprant was observed after 1, 6, and 50 mg/kg/day dosing regimen. Thecombined mean systemic exposure to grapiprant increased with dose in thedose range proportionally in the range 1-6 mg/kg/day, more thanproportionally in the range 6-50 mg/kg/day. Due to mild regeneration ofthe mucosal epithelium of the ileum in one male at 50 g/kg, the levelwith no observed adverse effects was 6 mg/kg. The combined mean C_(max)was 3,480 ng/mL and AUC₀₋₂₄ was 10,600 at 6 mg/kg. The combined meanC_(max) was 49,283 ng/mL and AUC₀-24 was 138,667 ng·h/mL.

There were no drug-related effects on mortality, bodyweight, foodconsumption, ophthalmology, electrocardiograms, hematology, coagulation,organ weights or gross pathological findings at doses up to 50 mg/kgadministered for a 9-month dosing period. Although functional effectssuch as loose or mucous stool and decreases in total protein, albuminand calcium in serum chemistry were observed at doses above 1 mg/kg,they were secondary gastrointestinal effects caused by the EP₄antagonism and, therefore, not adverse effects of the drug. Meanwhile,histopathological changes of mucosal epithelium in the ileum observed at50 mg/kg were considered an adverse effect of the drug.

Example 8—Effect of Grapiprant on Lameness and Pain in Dogs (CL-001)

Twenty intact adult female hounds underwent surgical meniscal release ofthe right stifle and were maintained for at least 8 weeks to allowosteoarthritis to develop. Radiographic signs of osteoarthritis wereconfirmed in all dogs about 8 weeks post-surgery. Animals wererandomized into three groups: negative control (n=6), positive control(n=7) treated with 5 mg/kg firocoxib daily, and grapiprant (n=7) dosedonce daily at 30 mg/kg. Dosing was based on baseline bodyweight.

Baseline physical exams, bodyweights, hematology, serum chemistry andurinalysis were obtained on study Day −1 and repeated on Day 13.Orthopedic assessments were conducted on Day −1, Day 2, Day 6, and Day13, including a kinetics assessment using the GAITRite system todetermine left hindlimb: right hindlimb ratios for peak pressure, stridelength, step length, and stance time, a 5-point scale lamenessevaluation, VAS assessments of right hindlimb function, right stiflepain and effusion, and comfortable range of motion measurements of bothstifles using a goniometer.

The groups did not differ significantly for any orthopedic parameterexcept for lower lameness scores in the grapiprant treated group on Day2 compared to the negative and positive control groups, and a higherfunction score (less lameness) in the negative control at baselinecompared to positive control and grapiprant treated groups. Thegrapiprant treated group and the negative control treated group both hadstatistically significant lower pain scores on Days 2 and 6 compared tobaseline. The positive control group improved statistically significantmean peak pressure ratios on Day 13 and in mean lameness and meanfunction scores on Day 6 and 13 compared to baseline. Grapiprant treatedanimals showed decreases in mean total protein, albumin, and globulin.

Grapiprant given at 30 mg/kg over 14 days was effective at day 2 inameliorating lameness and pain in the meniscal release model ofosteoarthritis. Grapiprant given at 30 mg/kg was considered safe for atleast a 14-day treatment period; however, serum protein levels decreasedslightly.

Example 9—Effectiveness of Grapiprant for Controlling Pain andInflammation in Feline Onychectomy (FCL-12-002)

This controlled, non-GLP, laboratory study used three treatment groups:Group 1 was dosed with 20-mg grapiprant-containing Formulation A34tablets the night before (about 12 hours) and 30 minutes before surgery;Group 2 was the negative control dosed with empty gelatin capsules; andGroup 3 was dosed with two 20-mg grapiprant-containing Formulation A34tablets given 30 minutes before surgery.

Ten adult cats were in each group. The cats were acclimated for 7 days.Cats were divided into two sequences of 15 cats each to allow forsurgeries over two days. The day of surgery was considered Day 0 forthat sequence. One cat from each grapiprant-treated group was removedbefore surgical procedures were performed. On Day −1/0 for thatsequence, all animals were administered with the appropriate testarticle before onychectomy. All animals received a dose of butorphanolbefore surgery. Following Day 2 procedures, all animals were transferredto an open feline colony. Study parameters of interest includedstatistical analysis of recovery, sedation, analgesia scores, andclinical pathology.

In summary, administering test article grapiprant was associated withdecreased analgesia scores (reduced pain) without an apparent effect onsedation scores compared to the placebo control. Both groups ofgrapiprant treated animals had fewer pain interventions than placebocontrol. Group 1 treatment regimen was associated with fewer paininterventions when compared to Group 3. Thus, dosing the cats withgrapiprant the night before surgery was more effective than providingthe same total amount of grapiprant to the cats only 30 minutes beforesurgery.

Example 10—Evaluating Dose Linearity and Effect of Feeding in Cats(FPK-11-001)

Grapiprant was blended in a mixture of excipients as shown in Table 14and administered orally by capsule to male and female cats. Animals wereassigned to groups as shown in Table 15.

TABLE 14 Grapiprant formulation used in this study Ingredient Component% (by weight) Grapiprant 45 Lactose 200 mesh 23 Dicalcium phosphatedihydrate 15 Pregelantized starch 5 Microcrystalline cellulose 6Povidone 5 Polaxamer 188 1

TABLE 15 Study groups. Number of Dose Level Group Animals (M/F) (mg/kg)Fed/Fasted 1 3/3 2 Fed 2 3/3 2 Fasted 3 3/3 6 Fasted 4 3/3 10 Fasted

The day of dosing was defined as Day 0. Following dosing, the animalswere evaluated for clinical signs. Blood samples were collected prior todosing and at 0.25, 0.5, 1, 2, 4, 8, 12, and 24-hours postdose. Theblood samples were processed to serum and analyzed for grapiprantconcentration. There were no test article-related clinical signs. Themean grapiprant pharmacokinetic results were as follow in Table 16.

TABLE 16 Dose Cmax Tmax AUC_(last) AUC_(0−∞) t_(1/2) Group (mg/kg) FoodSex (ng/mL) (hr) (hr*ng/mL) (hr*ng/mL) (hr) 1 2 Fed Male 407 1.65 10301220 3.12 1 2 Fed Female 327 1.33 990 1070 2.4 2 2 Fasted Male 1100 1.012290 1970 2.16 2 2 Fasted Female 1630 1.01 2780 2850 1.47 3 6 FastedMale 2970 0.99 3650 3750 2.79 3 6 Fasted Female 3110 1.17 6130 6350 4.984 10 Fasted Male 4280 1.33 7430 7580 4.03 4 10 Fasted Female 7130 0.7310700 10900 4.19

There were no sex differences although C_(max) and AUC values in femaleswere higher than in males for fasted cats, C_(max) and AUC were lower infemales for fed cats. Following a single oral dose of 2 mg/kg grapiprantcapsule to Group 1 fed cats, the mean T_(max) was 1.33 to 1.65 hours.The mean terminal half-life was 2.40 to 3.12 hours. Mean C_(max) was 327to 407 ng/mL, and mean AUC_(0-∞) was 1070 to 1220 hr*ng/mL.

After a single oral dose of 2 mg/kg grapiprant capsule to Group 2 fastedcats, the mean T_(max) was 1.01 hours. The mean terminal half-life was1.47 to 2.16 hours. Mean C_(max) was 1100 to 1630 ng/mL, and meanAUC_(0-∞) was 1970 to 2850 hr*ng/mL. Following an oral dose ofgrapiprant capsule at 2 mg/kg, C_(max) and AUCs were significantlyhigher (p<0.05) in fasted cats than in fed cats for both males andfemales. However, T_(max) and t_(1/2) were not significantly different.

After a single oral dose of 6 mg/kg grapiprant capsule to Group 3 fastedcats, the mean T_(max) was 0.99 to 1.17 hours. The mean terminalhalf-life was 2.79 to 4.98 hours. Mean C_(max) was 2970 to 3110 ng/mL,and mean AUC_(0-∞) was 3750 to 6350 hr*ng/mL.

After a single oral dose of 10 mg/kg grapiprant capsule to Group 4fasted cats, the mean T_(max) was 0.728 to 1.33 hours. The mean terminalhalf-life was 4.03 to 4.19 hours. Mean C_(max) was 4280 to 7130 ng/mL,and mean AUC_(0-∞) was 7580 to 10900 hr*ng/mL. Greater exposure occurredwhen animals were dosed in the fasted state than in the fed state, asevidenced by greater AUC and C_(max) for the Group 2 (fasted) than forthe Group 1 (fed) animals.

Exposure (AUC_(0-∞)) across groups of fasted animals (Groups 2-4) wasroughly dose-linear but less than dose-proportional. One-time grapiprantadministration (orally, via capsule) to male and female cats at doselevels of 2 (fed and fasted), 6, and 10 mg/kg was well-tolerated with notest article-related clinical signs. Significantly (p<0.05) greaterexposure occurred when animals were dosed in the fasted state than inthe fed state, as evidenced by greater AUC and C_(max) for the Group 2(fasted) than for the Group 1 (fed) animals. Exposure (AUC_(0-∞)) acrossgroups of fasted animals (Groups 2-4) was roughly dose-linear but lessthan dose-proportional.

Example 11—Evaluation of Binding of Grapiprant to Cat Serum Proteins byEquilibrium Dialysis (FPK-12-002)

Grapiprant concentrations of 200 and 1000 ng/mL in cat serum weresubjected to equilibrium dialysis using a Rapid Equilibrium Dialysis(RED) device. After dialysis, protein binding of grapiprant was high incat serum. The percent bound of grapiprant in cat serum at 200 and 1000ng/mL was 95% and 92%, respectively. Under these conditions, the changeof grapiprant concentration had no significant effect on cat serumprotein binding. The positive control compound warfarin (10 μM) was99.2% bound to human plasma proteins.

Example 12—Pharmacokinetics Grapiprant in Cats (FPK-12-003)

This controlled, non-GLP, laboratory study included two treatment groupsof six adult cats (three per sex) for a total of 12 study animals. Allcats underwent an 8-day acclimation phase. On Day 0, fasted animals inGroup 1 were orally-administered a 20-mg grapiprant-containing tablet(Formulation A34) while fasted animals in Group 2 wereorally-administered a 20-mg grapiprant-containing tablet (FormulationA29). Blood samples were collected and processed to serum forpharmacokinetic (PK) analysis. Four hours post-dose, all cats wereoffered food. Following PK sample collections on Day 1, all animals werereleased to an open feline colony. The variable of interest for thisstudy was the PK profile in serum. All animals were successfully dosedwith the designated test article formulation of grapiprant. No abnormalgeneral health was observed during the study. Grapiprant serumconcentrations were successfully measured to determine the PK profile asshown in Table 17.

TABLE 17 Cmax Tmax AUC_(last) AUC_(last)/Dose t_(1/2) Formulation SexStat (ng/mL) (hr) (hr*ng/mL) (hr*mg/mL) (hr) A29 Male Mean 1029 1.3 47601182 1.7 StDev 818 0.3 3564 832 0.4 CV % 79.5 43.3 74.9 — 25.3 FemaleMean 4413 1.2 17089 2905 1.7 StDev 1535 0.8 4189 1253 0.3 CV % 34.8 65.524.5 — 19.3 A34 Male Mean 1664 0.8 5785 1419 5.0 StDev 1090 0.3 2534 6382.9 CV % 65.5 34.6 43.8 — 57.9 Female Mean 9630 1.2 29691 4606 2.5 StDev4901 0.8 8780 968 0.7 CV % 50.9 65.5 29.6 — 27.7

In general, there were no abnormal general health observations duringthe study. Female cats demonstrated a greater systemic exposure thanmales for both the A29 and A34 formulations. Male cats demonstratedsimilar system exposure between the A29 and A34 formulations. Femalecats receiving grapiprant in the A34 formulation demonstrated greatersystemic grapiprant exposure than female cats administered the A29formulation. All female cats received a greater mg/kg dose than those ofmale cats; however, when adjusted for dose administered (mg/kg), femalecats continued to have a greater systemic grapiprant exposure than malecats.

Example 13—Pharmacokinetics of Grapiprant in Cats (FPK-13-004)

This controlled, non-GLP, laboratory study used three treatment groups:Group 1 was dosed with grapiprant at 2.5 mg/animal; Group 2 was dosedwith grapiprant at 5 mg/animal; and Group 3 was dosed with grapiprant at10 mg/animal. Each dose was administered once on Day 0. Blood sampleswere collected and processed to serum for pharmacokinetic (PK) analysis.Four hours post-dose, all cats were offered food. Following PK samplecollections on Day 1, all animals were released to an open felinecolony.

The mean pharmacokinetic parameters of grapiprant are presented in Table18 below. Pharmacokinetic parameters were similar between males andfemales within each dose group, and therefore mean values weredetermined by combining data from all animals.

TABLE 18 Pharmacokinetic Data. Dosage C_(max) C_(max)/Dose AUC_(last)AUC_(last)/Dose Dose (mg) (mg/kg) T_(max) (hr) (ng/mL) (kg*ng/mL/mg)(hr*ng/mL) (hr*ng/mL) t_(1/2) (hr) 2.5 0.5 1.50 1060 2120 4190 8380 1.585 1 1.13 2690 2500 10300 9450 1.13 10 2 1.25 4430 2150 21200 10300 3.44

Mean T_(max) values were similar among groups, ranging from 1.13 to 1.50hours. Elimination half-life values were similar between the 0.5 and 1mg/kg doses (1.58 and 1.13 hours, respectively) and increased slightlyfor the 2 mg/kg dose (3.44 hours). Increases in C_(max) were about doseproportional, and increases in AUC_(last) values were slightly greaterthan dose proportional. For a 4-fold increase in dose from 0.5 to 2mg/kg, C_(max) increased 4.18-fold, and AUC_(last) values increased5.06-fold. Overall, the data indicate that the 2 mg/kg dosage was themost effective in providing cats with grapiprant.

Example 14—Safety and Toxicokinetic Profiles of Grapiprant in Cats

The objective of this study was to evaluate the feline safety andtoxicokinetic profiles of grapiprant. Twenty-four domestic shorthaircats were randomly assigned to receive placebo or 3, 9, or 15 mg/kggrapiprant in a capsule formulation (3 males and 3 females per group)once daily for 28 days. All cats received their assigned medications asper protocol. Food consumption and behavior were observed daily,bodyweight measured weekly, and clinical pathology analyses on blood runon Days −7, 14, and 25, and urinalysis run on Days −7, and 25. Serialblood samples for toxicokinetic analyses were obtained following the Day0 and Day 27 doses. Full necropsies and histopathological evaluationswere performed following humane euthanasia on Day 28.

Grapiprant was well tolerated, and no adverse effects were noted atdaily doses up to 15 mg/kg for 28 days. All animals appeared normalthroughout the study. Grapiprant caused no deaths or drug-relatedeffects on body weight, food consumption, clinical pathology, organweight, gross pathology, or histopathology.

Grapiprant rapidly reached peak serum concentrations and maintainedsubstantial levels throughout the study. Grapiprant exposure varied withdose, but the 9 mg/kg and the 15 mg/kg groups showed similar meanAUC_(last) values. By Day 27, maximum grapiprant serum concentrationsranged from 683 ng/mL to 4950 ng/mL, and these were reached by 1-4 hoursafter administration. Mean half-lives on Day 27 ranged from about 3 toabout 14 hours, with a median value of about 5 to about 6 hours.

Grapiprant in a capsule formulation was well tolerated when given for 28days at serum exposures up to 4950 ng/mL.

Example 15—Further Pharmacokinetic Study of Grapiprant in Dogs

The objective of this study was to determine and compare thepharmacokinetic parameters of grapiprant in beagle dogs after singleoral administration of a flavored tablet formulation and amethylcellulose suspension formulation at nominal doses of 6 mg/kg and50 mg/kg. For this study, sixteen beagle dogs, two per sex per dose,were divided into two groups based on the nominal doses of grapiprant,either 6 mg/kg in Groups 3AB or 50 mg/kg in Groups 4A/B. Each group wasgiven the appropriate oral dose in a tablet or in a methylcelluloseformulation, using the crossover design as shown below at Table 19, onstudy Days 0 and 15. Blood samples were collected and serum wasprepared, frozen, and analyzed for concentration of grapiprant.

The vehicle was 0.5% methylcellulose (400 centipoise) in water. Abouthalf a volume of distilled water was heated to about 40-50° C.Methylcellulose was weighed to give a final concentration of 0.5% (w/v),and dissolved in the heated distilled water with stirring. The flaskwith the methylcellulose solution was cooled immediately. The remainingdistilled water was added into the flask to adjust volume. The finalsolution was stirred for least 30 minutes.

To prepare 1.2-mg/mL and 10-mg/mL suspensions grapiprant in the vehicle,an appropriate amount of grapiprant was added to a calibrated beakercontaining 0.5% aqueous methylcellulose, and mixed using a vortex. Thesuspension was homogenized using a Polytron™ mixer with attachedantifoam bit. A stir bar was added to the beaker, and the suspension wasstirred for about 5 minutes. The stir bar was removed temporarily toadjust the volume, and the suspension was stirred for another 5 minutesbefore it was transferred to a labeled amber dosing container or glassbeaker wrapped with foil.

The dosing suspension was prepared fresh on each day of dosing. A vortexdepth of between 75% and 100% of the depth of the container containingthe 500-mg suspension was achieved on both dosing days. Stirringsufficient enough to create a vortex of this depth resulted in ahomogenous mixture, as shown by all formulation samples having between93% and 100% of the nominal concentration. Dose formulation samples weretransported to the animal room before collection and were collected(while the formulations were being stirred) just before dosing the firstanimal of each group, and just before dosing the last animal of eachgroup. These steps guard against the 500-mg suspension settling duringtransport from the formulation laboratory to the animal room.

TABLE 19 Group Assignment and Dose Levels. No. of Group Animals Type ofDosing Day 0 Day 15 3A 2M/2F Tablet/Suspension 60-mg tablet 60-mgsuspension 3B 2M/2F Suspension/Tablet 60-mg suspension 60-mg tablet 4A2M/2F Tablet/Suspension Five 100-mg 500-mg tablets suspension 4B 2M/2FSuspension/Tablet 500-mg Five 100-mg suspension tablets

The gavage tube was closely examined postdose after dosing of allanimals on both days. No notable amounts of test article were observedin any gavage tube. There was no mortality. Slight increases in theincidence of fecal abnormalities were noted and are considered relatedto grapiprant exposure.

The T_(max) on Day 0 at the 60-mg dose was 1.0 or 2.0 hours for thetablet and 1.0 hour for the suspension. The average C_(max) for bothsexes combined was 5,330 ng/mL for the 60-mg tablet and 4,050 ng/mL forthe 60-mg suspension. The average AUC_(last) for both sexes combined was19,600 hr*ng/mL for the 60-mg tablet and 14,500 hr*ng/mL for the 60-mgsuspension.

The T_(max) on Day 0 at the 500-mg dose was 1.0, 2.0, or 4.0 hours forthe tablet and 0.5, 1.0, or 2.0 hours for the suspension. The averageC_(max) for both sexes combined was 96,100 ng/mL for the 500-mg tabletand 76,800 ng/mL for the 500-mg suspension. The average AUC_(last) forboth sexes combined was 455,000 hr*ng/mL for the 500-mg tablet and293,000 hr*ng/mL for the 500-mg suspension.

The T_(max) on Day 15 at the 60-mg dose was 1.0 or 2.0 hours for thetablet and 1.0 hour for the suspension. The average C_(max) for bothsexes combined was 6,110 ng/mL for the 60-mg tablet and 3,810 ng/mL forthe 60-mg suspension. The average AUC_(last) for both sexes combined was17,400 hr*ng/mL for the 60-mg tablet and 12,500 hr*ng/mL for the 60-mgsuspension.

The T_(max) on Day 15 at the 500-mg dose was 1.0 or 2.0 hours for thetablet and 1.0 hour for the suspension. The average C_(max) for bothsexes combined was 101,000 ng/mL for the 500-mg tablet and 74,200 ng/mLfor the 500-mg suspension. The average AUC_(last) for both sexescombined was 430,000 hr*ng/mL for the 500-mg tablet and 209,000 hr*ng/mLfor the 500-mg suspension.

T_(max), C_(max), and AUC_(last) were higher at the 500-mg dose than atthe 60-mg dose. C_(max) and AUC_(last) values were similar for a givendose on Days 0 and 15. There were differences in pharmacokineticparameters between the tablet and suspension at both doses, withexposure (AUC_(last)) greater in the tablet formulation versus thesuspension formulation. ANOVA indicated that for the tablet versus thesuspension formulations there was a significant difference (p≤0.05) forC_(max) and AUC_(last) at the 60-mg dose, and a significant difference(p≤0.05 or p≤0.01) for T_(max) and AUC_(last) at the 500-mg dose.Statistical differences occurred with and without the data from ananimal that vomited in the first 11 minutes of dosing. Overall, theAUC_(last) coefficients of variation of the 500-mg suspension were 53.7%(pharmacokinetic data from an animal with emesis excluded) on Day 0 and28.7% on Day 15.

Thus, results showed greater exposure (AUC) with the tablet formulationcompared to the suspension. The difference between AUCs for tablet andsuspension was statistically significant at both 60 mg and 500 mg, bothwhen animals with emesis were included and excluded from the analysis.The relative bioavailability ratio (tablet/suspension) for both doselevels, in both trials, was consistently greater than 100, whichindicated that tablet bioavailability was greater than suspensionbioavailability. The relative bioavailability ratio greater than 100difference persisted irrespective of whether animals with emesis wereincluded or excluded from the analysis.

What is claimed is:
 1. A method for treating pain and inflammation in acat or a dog, the method comprising administering to the cat or the dogan oral pharmaceutical composition comprising a therapeuticallyeffective amount of grapiprant and one or more excipients; wherein thetherapeutically effective amount of grapiprant is administered at adosage rate of 2 mg per kilogram bodyweight of the cat or the dog onceper day; and wherein the oral pharmaceutical composition is administeredwith or without food.
 2. The method of claim 1, wherein theadministering achieves a Cmax of grapiprant of 375 ng/mL to 10000 ng/mLat a Tmax of 0.4 to 3.4 hours.
 3. The method of claim 2, wherein theadministering achieves a Cmax of grapiprant of 750 ng/mL to 4000 ng/mL.4. The method of claim 3 wherein the administering achieves a Cmax ofgrapiprant of 1300 ng/mL to 4000 ng/mL.
 5. The method of claim 2,wherein the administering achieves the Cmax of grapiprant at a Tmax of0.7 to 1.7 hours.
 6. The method of claim 3, wherein the administeringachieves the Cmax of grapiprant at a Tmax of 0.5 to 1.0 hours.
 7. Themethod of claim 1, wherein the pharmaceutical composition furthercomprises 1% to 30% flavorant (w/w of the total composition).
 8. Themethod of claim 7, wherein the pharmaceutical composition comprises 5%to 15% flavorant (w/w of the total composition).
 9. The method of claim1, wherein the pharmaceutical composition is administered at least twicedaily.
 10. The method of claim 1, wherein the pharmaceutical compositionis administered for 6 days to 9 months.
 11. The method of claim 10,wherein the pharmaceutical composition is administered for 9 to 21 days.12. The method of claim 11, wherein the pharmaceutical composition isadministered for 12 to 14 days.
 13. The method of claim 1, wherein theadministering occurs from about 10 hours to about 18 hours before asurgery is performed on the cat or the dog.
 14. The method of claim 1,wherein the pharmaceutical composition comprises sodium lauryl sulfate.15. The method of claim 1, wherein the composition after administeringcauses no clinically significant adverse gastrointestinal event orchanges in liver, kidney, and coagulation parameters in the cat or thedog.